| fractionation of biologically active and inactive populations of human rhinovirus type 2. | | 1975 | 163526 |
| the effects of concanavalin a on the early events of infection by rhinovirus type 2 and poliovirus type 2. | the effect of concanavalin a (con a) on the course of early infection of hela cells with purified radioactive human rhinovirus type 2 (hrv-2) or poliovirus type 2 (p-2) has been examined. several early steps in infection were inhibited before the uncoating of parental virus. con a, at 100 mug/ml, reduces attachment of virus when added to cells before infection. con a also detectably slows the normal progression of adsorbed virus to tightly bound forms characterized, in the case of hrv-2, by resi ... | 1975 | 170375 |
| inhibition by zinc of rhinovirus protein cleavage: interaction of zinc with capsid polypeptides. | zinic ions rapidly inhibit virus production in hela cells infected with human rhinovirus type 1a and lead to the accumulation of human rhinovirus type 1a precursor polypeptides. the degree to which cleavage of these precursors is inhibited is directly dependent on the quantity of cell-associated zinc. proteolysis resumes after the removal of zinc-containing medium, and the accumulated viral precursors are cleaved predominantly to stable virus polypeptides. the precursors stabilized at the lowest ... | 1976 | 176466 |
| physical and metabolic requirements for early interaction of poliovirus and human rhinovirus with hela cells. | attachment, ""tight binding'' and eclipse of radioactive poliovirus 2 (p2) and human rhinovirus 2 (hrv 2) were investigated. the activation energy for attachment of both hrv2 and p2 was about 13 kcal/mol. hrv2 differed from p2 in two respects: the arrhenius plot for attachment of hrv2 showed a break at 15 to 19 degrees c when the cells were first treated several hours at 0 degrees c, and attachment of hrv2 was inhibited by treatment of cells with metabolic poisons able to reduce cellular atp by ... | 1976 | 184301 |
| polyadenylate sequences of human rhinovirus and poliovirus rna and cordycepin sensitivity of virus replication. | the polyadenylate [poly(a)] content of the genome rna of human rhinovirus type 14 (hrv-14) is nearly twice as large as that of the genome rna of poliovirus type 2. the poly(a) content of viral rna was determined to be the rnase-resistant fraction of 32p-labeled viral rna extracted from purified virions. polyacrylamide gel electrophoresis indicated that the poly(a) sequences of hrv-14 are more heterogenous and on an average larger than those of poliovirus rna. on the basis of susceptibility to mi ... | 1976 | 185411 |
| gel double immunodiffusion studies with sex human rhinoviruses. | rabbits were immunized on six occasions during a seven month period with fluorocarbon and sucrose denstiy gradient purified preparations of human rhinovirus types 1a,2,3,4,9, or 14. sera collected 1 week after the final immunization formed 1 or 2 preciptin lines when reacted by immunodiffusion against fluorocarbon purified preparations of each homologous immunizing virus. heterologous preciptin cross reactions were detected between: rv1a antigen and rabbit sera to rv2, rv9 and rv14; rv2 antigen ... | 1978 | 209768 |
| picornaviruses: rapid differentiation and identification by immune electronmicroscopy and immunodiffusion. | immune electronmicroscopy (iem) was used to identify human picornaviruses rapidly and to differentiate enteroviruses from rhinoviruses. human sera, diluted 10- to 50-fold beyond the neutralisation endpoints for homologous virus, readily agglutinated c-type antigens of seven human picornaviruses. human sera did not react by iem with a control animal picornavirus. by iem after acid treatment, differentiation of a human enterovirus from a human rhinovirus was possible. there was an excellent correl ... | 1977 | 404426 |
| identification of 50- and 23-/25-kda hela cell membrane glycoproteins involved in poliovirus infection: occurrence of poliovirus specific binding sites on susceptible and nonsusceptible cells. | glycoproteins in the range 50 and 23/25 kda were identified as poliovirus specific binding sites on hela cells with the monoclonal antibody mab 122. mab 122 is characterized by its partial inhibiting effect on poliovirus reproduction and adsorption when prebound to hela cells. the binding sites are endocytosed in native cells and specific for poliovirus as mab 122 did not interfere with the adsorption of human rhinovirus type 14 (hrv 14). the poliovirus binding sites are present also on nonprima ... | 1992 | 1310182 |
| comfa analysis of the interactions of antipicornavirus compounds in the binding pocket of human rhinovirus-14. | a comfa analysis of eight compounds related to disoxaril whose x-ray structures bound to hrv-14 had been determined resulted in a strong positive correlation of activity with steric effects of the compounds, particularly toward the pore end of the compound binding site, and no correlation with electrostatic effects. these results confirm what had been previously found, that the activity of these compounds was highly dependent upon their hydrophobic nature as expressed by log p. the comfa study a ... | 1992 | 1313108 |
| initiation of translation of human rhinovirus rna: mapping the internal ribosome entry site. | in order to map the 3' boundary of the segments needed for translation initiation at the correct site on human rhinovirus 2, deletions were made from the 3' end of the viral 5'-untranslated region. these truncated viral segments were placed immediately upstream of a reporter gene, a derivative of the influenza virus ns cdna, either as monocistronic constructs or as dicistronic constructs in which the upstream cistron was the xenopus laevis cyclin b2 cdna. in vitro transcripts of these clones wer ... | 1992 | 1316679 |
| in vitro activity of pirodavir (r 77975), a substituted phenoxy-pyridazinamine with broad-spectrum antipicornaviral activity. | pirodavir (r 77975) is the prototype of a novel class of broad-spectrum antipicornavirus compounds. although its predecessor, r 61837, a substituted phenyl-pyridazinamine, was effective in inhibiting 80% of 100 serotypes tested (ec80) at concentrations above 32 micrograms/ml, pirodavir inhibits the same percentage of viruses at 0.064 micrograms/ml. whereas r 61837 was active almost exclusively against rhinovirus serotypes of antiviral group b, pirodavir is broad spectrum in that it is highly act ... | 1992 | 1317142 |
| binding of an antiviral agent to a sensitive and a resistant human rhinovirus. computer simulation studies with sampling of amino acid side-chain conformation. i. mapping the rotamers of residue 188 of viral protein 1. | the mutation of valine 188 to leucine in the viral protein 1 of human rhinovirus 14 renders the virus resistant to certain antiviral compounds. thermodynamic-cycle perturbation theory provides a means of calculating the difference in the binding free energies of an antiviral compound to the wild-type virus and to the mutant virus. in calculating the relevant free-energy differences in molecular dynamics simulations, it is important to sample the multiple rotational isomers of residue 188 correct ... | 1992 | 1318383 |
| binding of an antiviral agent to a sensitive and a resistant human rhinovirus. computer simulation studies with sampling of amino acid side-chain conformations. ii. calculation of free-energy differences by thermodynamic integration. | thermodynamic-cycle perturbation theory and molecular dynamics simulations were used to calculate the difference in the free energy of binding of the antiviral compound win53338 to the wild-type human rhinovirus 14 and to a drug-resistant mutant of the virus in which valine 188 of the viral protein 1 is mutated to leucine. because of the difficulty of achieving adequate sampling of all of the rotational isomers of amino acid side-chains in molecular dynamics simulations, an explicit treatment of ... | 1992 | 1318384 |
| study of the parameters of binding of r 61837 to human rhinovirus 9 and immunobiochemical evidence of capsid-stabilizing activity of the compound. | the binding of the antiviral compound r 61837 to human rhinovirus 9 (hrv 9) was studied quantitatively and compared with binding of r 61837 to hrv 9h, a semiresistant variant. for both strains, radiolabelled r 61387 bound to native particles only. the kd values obtained by scatchard analysis of saturation binding data were 37 nm for hrv 9 and 172 nm for hrv 9h, whereas the concentrations resulting in a 50% reduction of cytopathic effect were 42 nm and 840 nm, respectively. reversibility experime ... | 1992 | 1318682 |
| using proteinase trapping to detect revertants of inactive rhinoviral 2a proteinase mutants. | the 2a proteinase of human rhinovirus 2 cleaves itself off the growing polyprotein at its own n terminus during translation; this property was used to develop an in vivo screening system with the lacz gene fragment of m13mp18. the fusion of an active 2a proteinase to the c-terminus of the alpha-fragment did not affect alpha-complementation, as the proteinase cleaved itself off the alpha-fragment. however, an inactive 2a proteinase remained fused to the alpha-fragment hindering alpha-complementat ... | 1992 | 1325156 |
| cleavage specificity on synthetic peptide substrates of human rhinovirus 2 proteinase 2a. | proteinase 2a of human rhinovirus serotype 2 (hrv2 2a) was expressed in escherichia coli and partially purified; the preparation was used to study various enzymatic parameters. using a 16-amino acid peptide representing the native cleavage region of hrv2 2a, an apparent km value of 5.4 x 10(-4) mol/liter was determined. a minimum of 9 amino acids (comprising residues p8 to p1') was necessary for cleavage to occur. proteolysis of substituted peptides was highly tolerant toward changes at p1, p2', ... | 1992 | 1331062 |
| acid-induced structural changes in human rhinovirus 14: possible role in uncoating. | x-ray diffraction data were collected from human rhinovirus 14 crystals a few minutes after exposure to acid vapor and prior to excessive crystalline disorder. conformational changes occurred (i) in the gh loop of viral protein (vp) 1, (ii) at the ion binding site on the outer surface of the pentamer center, and (iii) in vp3 and vp4 on the virion's interior in the vicinity of the fivefold axis. amino acid substitutions in mutants resistant to low ph, or to drugs that inhibit uncoating, were conc ... | 1992 | 1332036 |
| purification and crystallization of intact human rhinovirus complexed with a neutralizing fab. | we report the first crystallization of an intact virion, human rhinovirus 14, complexed with the fab fragment from a neutralizing antibody. these crystals diffract to at least 6.0a resolution. it has been suggested that fab's and mab's can induce large conformational changes in the capsid upon binding. the structure of this complex should enable us to detect the existence and role of such changes. | 1992 | 1333115 |
| antirhinoviral activity of heterocyclic analogs of win 54954. | a variety of heterocyclic analogs of win 54954 have been synthesized and tested in vitro against human rhinovirus type 14 (hrv-14) in a plaque reduction assay. the more active compounds were tested against 14 additional serotypes, and the concentration which inhibited 80% of the serotypes tested (mic80) was measured. one compound, 37, exhibited activity comparable to win 59454. physicochemical as well as electrostatic parameters were calculated and the results subjected to a qsar analysis in an ... | 1992 | 1335079 |
| conformationally restricted analogues of disoxaril: a comparison of the activity against human rhinovirus types 14 and 1a. | a series of conformationally restricted analogs of disoxaril has been synthesized and evaluated against human rhinovirus types (hrv) 14 and 1a. the sensitivity of these serotypes to this series varied and was dependent upon the length of the molecule as well as upon the flexibility of the aliphatic chain. minimum energy conformations of these compounds were overlaid with the x-ray structure of a closely related analog 9 bound to the capsid protein of both hrv-14 and -1a and then modeled in the c ... | 1992 | 1335081 |
| three-dimensional structure of the fab fragment of a neutralizing antibody to human rhinovirus serotype 2. | the crystal structure of the antigen-binding fragment of a monoclonal antibody (8f5) that neutralizes human rhinovirus serotype 2 has been determined by x-ray diffraction studies. antibody 8f5, obtained by immunization with native hrv2 virions, cross-reacts with peptides of the viral capsid protein vp2, which contribute to the neutralizing immunogenic site b in this serotype. the structure was solved by the molecular replacement method and has been refined to an r-factor of 18.9% at 2.8 a resolu ... | 1992 | 1338980 |
| plasmodium falciparum-infected erythrocytes bind icam-1 at a site distinct from lfa-1, mac-1, and human rhinovirus. | the attachment of erythrocytes infected with p. falciparum to human venular endothelium is the primary step leading to complications from severe and cerebral malaria. intercellular adhesion molecule-1 (icam-1, cd54) has been implicated as a cytoadhesion receptor for p. falciparum-infected erythrocytes. characterization of domain deletion, human/murine chimeric icam-1 molecules, and amino acid substitution mutants localized the primary binding site for parasitized erythrocytes to the first amino- ... | 1992 | 1346257 |
| regulation of icam-1 expression and function in human dermal fibroblasts by il-4. | icam-1 is found on the surface of many hematopoietic and nonhematopoietic cells and can function as an adhesive ligand for the integrin, leukocyte function-associated molecule-1 (lfa-1, cd11a/cd18). icam-1/leukocyte function-associated molecule-1 interaction has been shown to be of importance in many immune-mediated cell-cell adhesion reactions. in vitro, unstimulated human fibroblast cell cultures express low levels of icam-1. using elisa, cytofluorography, electron microscopy, northern analysi ... | 1992 | 1347050 |
| internalization of a major group human rhinovirus does not require cytoplasmic or transmembrane domains of icam-1. | intercellular adhesion molecule-1 (cd54), a cell adhesion molecule and the receptor for the major group of rhinoviruses, is a class 1 membrane protein with five ig-like domains in its extracellular region, a transmembrane domain, and a short cytoplasmic domain. the amino-terminal domains (d1 and d2) are sufficient for virus binding and the first is most important (1). we have investigated whether other extracellular domains, transmembrane or cytoplasmic domains are required for virus entry as de ... | 1992 | 1349619 |
| preliminary x-ray crystallographic analysis of intercellular adhesion molecule-1. | crystals of the two amino-terminal domains of intercellular adhesion molecule-1, the receptor for the major group of human rhinovirus serotypes, diffract to 3.0 a resolution. the crystals are trigonal in space group p3(1)21 or p3(2)21 with cell dimensions of a = b = 55.7 a, c = 166.3 a, with probably six molecules per unit cell. | 1992 | 1351949 |
| in vitro studies of the antirhinovirus activity of soluble intercellular adhesion molecule-1. | we studied the in vitro antirhinovirus activity of a soluble form of intercellular adhesion molecule-1 (sicam-1). sicam-1 inhibited the cytopathic effect of 10 representative human rhinovirus (hrv) serotypes of the major receptor group with, 50% effective concentrations (ec50s) of 0.1 to 7.9 micrograms/ml. cell type-dependent variation in the inhibitory activity of sicam-1 was observed for two major receptor group serotypes in hela cells (ec50, greater than 32 micrograms/ml), and no inhibitory e ... | 1992 | 1358025 |
| ion channels in icosahedral virus: a comparative analysis of the structures and binding sites at their fivefold axes. | an analysis of the crystallographically determined structures of the icosahedral protein coats of tomato bushy stunt virus, southern bean mosaic virus, satellite tobacco necrosis virus, human rhinovirus 14 and mengovirus around their fivefold axes is presented. accessibilities surfaces, electrostatic energy profile calculations, ion-protein interaction energy calculations, free energy perturbation methods and comparisons with structures of chelating agents are used in this study. it is concluded ... | 1992 | 1384743 |
| in vitro virucidal effects of allium sativum (garlic) extract and compounds. | garlic (allium sativum) has been shown to have antiviral activity, but the compounds responsible have not been identified. using direct pre-infection incubation assays, we determined the in vitro virucidal effects of fresh garlic extract, its polar fraction, and the following garlic associated compounds: diallyl thiosulfinate (allicin), allyl methyl thiosulfinate, methyl allyl thiosulfinate, ajoene, alliin, deoxyalliin, diallyl disulfide, and diallyl trisulfide. activity was determined against s ... | 1992 | 1470664 |
| conservation of rna-protein interactions among picornaviruses. | picornavirus genomes encode unique 5' noncoding regions (5' ncrs) which are approximately 600 to 1,300 nucleotides in length, contain multiple upstream aug codons, and display the ability to form extensive secondary structures. a number of recent reports have shown that picornavirus 5' ncrs are able to facilitate cap-independent internal initiation of translation. this mechanism of translation occurs in the absence of viral gene products, suggesting that the host cell contains the necessary comp ... | 1992 | 1602550 |
| proteinase trapping: screening for viral proteinase mutants by alpha complementation. | many virally encoded proteinases cleave themselves out of a polyprotein, with cleavage occurring usually at their own n terminus. this property was used to develop an in vivo screening system using the lacz gene fragment of m13mp18. when a fusion protein of the alpha fragment of beta-galactosidase and an active 2a proteinase of human rhinovirus 2 was expressed, alpha complementation was not affected, as the 2a proteinase cleaved itself off the alpha fragment. however, fusion of an inactive 2a pr ... | 1991 | 1648726 |
| human rhinovirus mutants resistant to low ph. | mutants of human rhinovirus serotype 14 (hrv14) with increased resistance to treatment at low ph were obtained by repeated cycles of exposure to ph 4.5 and propagation in hela cells. whereas wild-type virus lost more than 5 logs of infectivity upon incubation at ph 4.3, the three isolates examined were essentially unaffected. conformational change of the viral capsid upon exposure to low ph was assessed as an increase of hydrophobicity by partition between an aqueous phase and a triton x-114 pha ... | 1991 | 1649506 |
| localization of rhinovirus replication in vitro with in situ hybridization. | to facilitate understanding of human rhinovirus (hrv) pathogenesis, methods were developed for detection of hrv infection in vitro using in situ hybridization (ish). hrv-14 rna probes and oligonucleotide probes representing conserved sequences in the 5'-non-translated region were labeled with 35s and used to detect infected hela or wi-38 strain human embryonic lung cells in cytological preparations. ish was shown to be specific for detection of hrv on a single-cell basis. subsequently, in human ... | 1991 | 1653307 |
| biologically active protease 3c of human rhinovirus 1a is expressed from a cloned cdna segment in escherichia coli. | we produced the putative protease 3c of human rhinovirus 1a (hrv-1a), a minor group rhinovirus, by escherichia coli expression of a segment of hrv-1a cdna coding for 3a, 3b, 3c, and parts of 2c and 3d (delta 2c3abc delta 3d). the protease 3c was expected to be processed by intramolecular q-g cleavages from the virus-specific precursor polypeptide. while the n-terminal 3b-3c site was correctly cleaved, the c-terminal q-g site of 3c was not processed. western blotting with a site-specific polyclon ... | 1991 | 1653490 |
| construction and characterization of a poliovirus/rhinovirus antigenic hybrid. | in order to study the properties of foreign antigenic sites expressed on poliovirus a hybrid was constructed in which neutralization antigenic site ia of poliovirus type 1 strain mahoney [pv1(m)] was replaced by neutralization immunogenic site ia (nimia) of human rhinovirus 14 (hrv14). the resulting hybrid was viable, but growth was impaired in comparison to pv1(m). the hybrid expressed both pv1(m) and hrv14 antigenic determinants. when inoculated into rabbits it elicited neutralizing antibodies ... | 1991 | 1653493 |
| development of synthetic peptide substrates for the poliovirus 3c proteinase. | picornaviruses, such as polio, translate their entire genome as a single polyprotein which must be proteolytically processed to produce the mature viral proteins. a majority of these cleavages are catalyzed by the virus-encoded cysteine proteinase, 3c. we report here the design and synthesis of a series of oligopeptide substrates, based upon native 3c cleavage sites, for an hplc assay of poliovirus 3c proteinase activity. a similar series of peptides based upon human rhinovirus 3c cleavage sites ... | 1991 | 1654789 |
| binding affinities of structurally related human rhinovirus capsid-binding compounds are related to their activities against human rhinovirus type 14. | the binding affinities (kds) and the rates of association and dissociation of members of a chemical class of antiviral compounds at their active sites in human rhinovirus type 14 (hrv-14) were determined. on the basis of analysis by ligand, a nonlinear curve-fitting program, of saturation binding experiments with hrv-14, the kds for win 52084, win 56590, disoxaril (win 51711), and win 54954 were found to be 0.02, 0.02, 0.08, and 0.22 microm, respectively. the independently determined kinetic rat ... | 1991 | 1656851 |
| rhinovirus 3c protease catalyzes efficient cleavage of a fluorescein-labeled peptide affording a rapid and robust assay. | the 3c protease encoded by human rhinovirus type 2 catalyzes with equal efficiency cleavage of a peptide substrate with or without a fluorescein label attached to the amino acid at the p7' position. substrates ac-mealfqgplqykdl-nh2 and mealfqgplqyke(fluorescein)l are hydrolyzed with values of vmax/km of 970 m-1 s-1 and 1100 m-1 s-1, respectively. with the labeled substrate, hplc achieves separation of substrate and product in 2.5 min. separation in as little as 12 s is feasible. fluorescein was ... | 1991 | 1658106 |
| rhinovirus induces natural killer-like cytotoxic cells and interferon alpha in mononuclear leukocytes. | natural killer-like cellular cytotoxicity was augmented by incubation of human rhinovirus serotype 2 with peripheral blood mononuclear leukocytes collected from healthy donors. the production of alpha interferon but not gamma interferon was identified in the same cell cultures. a specific interaction of conformationally intact rhinovirus with peripheral blood mononuclear leukocytes was required for induction of the response, since the response was extinguished at reduced quantities of infectious ... | 1991 | 1662702 |
| in vitro virucidal activity of selected anthraquinones and anthraquinone derivatives. | anthraquinones and anthraquinone derivatives were characterized for their antiviral and virucidal activities against viruses representing several taxonomic groups. one of these compounds, hypericin, had activity against vesicular stomatitis virus, herpes simplex virus types 1 and 2, parainfluenza virus, and vaccinia virus (from 0.5 to 3.8 log10 reductions in infectivity) at concentrations of less than 1 microgram/ml as determined by a direct pre-infection incubation assay. human rhinovirus was n ... | 1991 | 1665961 |
| a comparative test of fifteen compounds against all known human rhinovirus serotypes as a basis for a more rational screening program. | a systematic evaluation of 15 rhinovirus capsid-binding agents against all 100 serotyped human rhinoviruses revealed the existence of two virus groups, based upon differential susceptibility to antiviral compounds. elongated and short-chained compounds preferentially inhibited groups a and b. the positions of the rhinoviruses within a map derived from a multivariate analysis allow for the selection of a panel of 17 rhinoviruses, for which the median antiviral inhibitory value against them will a ... | 1991 | 1666824 |
| antiviral activity of constituents of tamus communis. | the antiviral activity of the phenanthrene derivatives 1-6, of the spyrostane triglycosides dioscin (7) and gracillin (8), of the furostanol tetraglycosides methylprotodioscin (9), its (25s) epimer methylprotoneodioscin (10), and methylprotogracillin 11, have been tested towards two rna viruses: vesicular stomatitis virus and human rhinovirus type 1b. all these products were extracted from the rizomes of tamus communis l; compound 11 was isolated also from asparagus cochinchinesis, together with ... | 1991 | 1667189 |
| enantiomeric separation of substituted flavanoids by lc-dad. | a variety of racemic flavanoids with anti-rhinovirus activity have been resolved for the first time by hplc, using a chiral stationary phase. baseline separation was easily obtained for racemic 4',6-dichloroisoflavan (v). the absolute configurations of two enantiomers (va and vb) were established by comparing their circular dichroism curves with those of other known isoflavans. both the isomers were tested against human rhinovirus serotype 1b infection in vitro; the s form was approximately four ... | 1991 | 1668301 |
| poliovirus can enter and infect mammalian cells by way of an intercellular adhesion molecule 1 pathway. | mouse fibroblast cell lines were transfected with truncated forms of the human poliovirus receptor (pvr) cdna and tested for the expression of functional receptors for poliovirus. several receptor constructs, all containing the coding region of the first 143 amino acids of pvr, were able to render mouse cells susceptible to poliovirus infection. a deletion of 65 amino acids in the first extracellular domain of pvr prevented virus attachment and infection. these data suggest that domain 1 is nece ... | 1991 | 1673787 |
| binding of human rhinovirus and t cells to intercellular adhesion molecule-1 on human fibroblasts. discordance between effects of il-1 and ifn-gamma. | intercellular adhesion molecule-1 (icam-1) is found on the surface of many hemopoietic and non-hemopoietic cells and can function as an adhesive ligand for the integrin, leukocyte function associated molecule-1 (lfa-1, cd11a/cd18). icam-1/lfa-1 interaction is thought to be of importance in many immune mediated cell-cell adhesion reactions. recently, the major human rhinovirus (hrv) receptor has been identified as icam-1. hrv has been shown to bind specifically to icam-1 on transfected cos cells ... | 1991 | 1679837 |
| neutralization epitopes of human rhinovirus type 2. | fourteen neutralizing monoclonal antibodies recognizing human rhinovirus (hrv) type 2 have been used to select a total of 51 virus escape mutants. cross-resistance analysis of the mutants, together with rna sequencing and identification of amino acid substitutions, have revealed three neutralization sites on the virus surface. two of these appear to correspond to the nim-ia and nim-ii sites described for hrv-type 14, although there are also substantial differences. the third site has not been de ... | 1990 | 1693662 |
| structural and serological evidence for a novel mechanism of antigenic variation in foot-and-mouth disease virus. | changes resulting in altered antigenic properties of viruses nearly always occur on their surface and have been attributed to the substitution of residues directly involved in binding antibody. to investigate the mechanism of antigenic variation in foot-and-mouth disease virus (fmdv), variants that escape neutralization by a monoclonal antibody have been compared crystallographically and serologically with parental virus. fmdvs form one of the four genera of the picornaviridae. the unenveloped i ... | 1990 | 1699132 |
| neutralizing antibodies to human rhinovirus produced in laboratory animals and humans that recognize a linear sequence from vp2. | synthetic peptides representing amino acids 156 to 170 of virus capsid protein vp2 from human rhinovirus (hrv) type 2 have previously been used to elicit a neutralizing antibody response; polyclonal antisera against this peptide have been compared with a virus-neutralizing monoclonal antibody (8f5) which recognizes the same linear sequence. the neutralizing activity of both antibodies against virus is abolished by an amino acid substitution in vp2 at position 163 but only the peptide antiserum n ... | 1990 | 1703215 |
| characterization of mengo virus neutralization epitopes. | a set of four monoclonal antibodies which neutralized the infectivity of mengo virus was used to select 20 non-neutralizable (escape) mutants. altered amino acids were identified by sequence analyses of the capsid-coding regions of the mutant virus genomes. mutations were found predominantly in proteins vp2 and vp3, while mutations in vp1 were detected only as second mutations. the mengo virus vp2 mutations at amino acid residues 2144, 2145, 2147, and 2148 align with site nlm ii in human rhinovi ... | 1991 | 1704653 |
| proliferative responses of t cells primed against human rhinovirus to other rhinovirus serotypes. | lymphocytes from mice immunized with human rhinovirus (hrv) serotypes 1a or 15 proliferated in vitro in response to hrv and the activated cells were shown to be helper t (th) cells. lymphocytes from mice primed with hrv-1a responded to seven of eight heterologous virus serotypes, the responses to other minor cell receptor group viruses being greater than to those belonging to the major cell receptor group. a similar bias was seen with cells from mice primed with hrv-15 in that they responded pre ... | 1991 | 1722503 |
| foreign epitopes in immunodominant regions of hepatitis b core particles are highly immunogenic and conformationally restricted. | the presentation of heterologous amino acid sequences on the surface of hepatitis b core antigen (hbcag) particles has been studied using a defined linear neutralization site from human rhinovirus (hrv). previous work has shown that fusion particles, in which the hrv peptide sequence is linked to the amino terminus of the hbcag protein, induce excellent immune responses in experimental animals. using predictive models of hbcag particulate structure and the approximate location of the major immun ... | 1991 | 1722937 |
| the major and minor group receptor families contain all but one human rhinovirus serotype. | previous studies have assigned 88 human rhinovirus (hrv) serotypes to major and minor receptor groups. extension of these studies to include the remaining 14 unassigned serotypes indicated that 13 serotypes belong to the major group since their infection of hela cells is completely blocked by a monoclonal antibody that recognizes the major group receptor. this result indicates that the major group now accounts for 91 of the 102 known serotypes, while the minor group contains 10 serotypes. one se ... | 1991 | 1846502 |
| stabilization of human rhinovirus serotype 2 against ph-induced conformational change by antiviral compounds. | four win compounds with anti-picornavirus activities were tested for their ability to stabilize human rhinovirus serotype 2 (hrv-2) against low ph-induced conformational changes in vitro, as determined by specific immunoprecipitation. these results were compared to the minimal inhibitory concentration (mic) as measured in a plaque reduction assay. a direct relationship was observed between the concentration of the compound that prevented the low ph-induced conformational changes and the mic, ind ... | 1991 | 1847178 |
| human rhinovirus 14 complexed with antiviral compound r 61837. | the binding of the antirhinoviral agent r 61837 to human rhinovirus 14 has been examined by x-ray crystallographic methods. the compound r 61837 binds in the same pocket (underneath the canyon floor) as the "win" antirhinoviral agents. it does not penetrate as far into the pocket but causes similar conformational changes in the virus capsid. the movement of residues 1217 to 1221 of viral protein 1 (in the "fmdv loop") is more pronounced for r 61837 than for win compounds. although both r 61837 a ... | 1991 | 1847215 |
| substrate requirements of a human rhinoviral 2a proteinase. | the genetic information contained within the rna genome of picornaviruses is expressed as a single large open reading frame; processing of the primary translation product begins while translation is still in progress. in rhinoviruses and enteroviruses, two picornavirus genera, the virally encoded proteinase 2a begins the processing cascade, cleaving between the c-terminus of vp1 and its own n-terminus. the natural variation in the amino acid sequences amongst rhinoviruses and enteroviruses at th ... | 1991 | 1847268 |
| detection of rhinovirus rna in nasal epithelial cells by in situ hybridization. | this paper describes the development and evaluation of in situ hybridization (ish) for the detection of rhinovirus in cells obtained from nasal washings of volunteers infected with human rhinovirus 14 (hrv-14). twenty-five (66%) and 27 (71%) of 38 volunteers inoculated with hrv-14 had evidence of infection by virus isolation and ish, respectively, on at least one of 4 days investigated after virus challenge. in contrast, only 14 of 38 (37%) volunteers had significant antibody rises as detected b ... | 1990 | 1964938 |
| functional aspects of the capsid structure of mengo virus. | the three-dimensional structure of the mengo virus capsid has been determined at a resolution of 3.0 a. this achievement is discussed in an historical context, and the general features of picornavirus capsid design are presented. the dynamic functional aspects of the mengo virus capsid--namely its ability to interact with specific receptors on host cells, to dissociate and release the viral genomic rna into the cellular cytoplasm, to assemble with progeny rna molecules and form new virions, and ... | 1990 | 1965133 |
| antibodies that block rhinovirus attachment map to domain 1 of the major group receptor. | the vast majority of human rhinovirus serotypes utilize the intercellular adhesion molecule 1 (icam-1) as the attachment site on susceptible cells. twelve murine monoclonal antibodies were isolated and shown by competition binding studies to recognize three distinct, nonoverlapping epitopes on the icam-1 receptor. titration of three antibodies representing each of the binding sites demonstrated that they were equally effective at blocking viral attachment. by using in vitro transcription and tra ... | 1990 | 1970837 |
| modeling of the human intercellular adhesion molecule-1, the human rhinovirus major group receptor. | a model has been built of the amino-terminal domain of the intercellular adhesion molecule-1 (icam-1), the receptor for most human rhinovirus serotypes. the model was based on sequence and presumed structural homology to immunoglobulin constant domains. it fits well into the putative receptor attachment site, the canyon, on the human rhinovirus-14 (hrv14) surface in a manner consistent with most of the mutational data for icam-1 (staunton, d. e., dustin, m. l., erickson, h. p., springer, t. a. c ... | 1990 | 1972986 |
| bacterial expression of antibody fragments that block human rhinovirus infection of cultured cells. | human rhinoviruses are the major causative agents of the common cold in humans and have been divided into major and minor groups based on receptor specificity. cdnas encoding the light and heavy chains of a murine monoclonal antibody that recognizes the major group receptor were cloned, abundantly expressed in escherichia coli, and renatured into fab fragments that blocked virus binding and protected hela cell monolayers from rhinovirus infection. elimination of the cysteines normally bridging t ... | 1990 | 2153680 |
| limited expression of poliovirus by vaccinia virus recombinants due to inhibition of the vector by proteinase 2a. | a recombinant vaccinia virus was constructed that expressed poliovirus coat precursor protein p1 fused to about two-thirds of the 2a proteinase. the truncated 2a segment could be cleaved away from the p1 region by coinfecting with poliovirus type 1, 2, or 3 or with human rhinovirus 14 but not with encephalomyocarditis virus. further cleavage of the vector-derived p1 to yield mature poliovirus capsid proteins was not observed. attempts to isolate vaccinia virus recombinants containing portions of ... | 1990 | 2154618 |
| analysis of the structure of a common cold virus, human rhinovirus 14, refined at a resolution of 3.0 a. | human rhinovirus 14 has a pseudo t = 3 icosahedral structure in which 60 copies of the three larger capsid proteins vp1, vp2 and vp3 are arranged in an icosahedral surface lattice, reminiscent of t = 3 viruses such as tomato bushy stunt virus and southern bean mosaic virus. the overall secondary and tertiary structures of vp1, vp2 and vp3 are very similar. the structure of human rhinovirus 14, which was refined at a resolution of 3.0 a [r = 0.16 for reflections with f greater than 3 sigma(f)], i ... | 1990 | 2156077 |
| structural refinement and analysis of mengo virus. | the structure of mengo encephalomyelitis virus was refined at 3 a resolution with a final r-factor of 0.221 and a root-mean-square deviation from idealized bond lengths of 0.019 a for 10 a to 3 a data with f greater than or equal to 3 sigma(f). the hendrickson-konnert refinement was restrained by the phases derived from the molecular replacement averaging procedure and constrained by the icosahedral symmetry of the virus. the virus consists of 60 protomers each having three major subunits, vp1, ... | 1990 | 2156078 |
| a model for compounds active against human rhinovirus-14 based on x-ray crystallography data. | a number of (oxazolinylphenyl)isoxazoles have been synthesized and tested against human rhinovirus-14 (hrv-14). several of the more active compounds have been examined by x-ray crystallography and their orientation in the compound binding site on the capsid protein of hrv-14 has been determined. based on the minimum inhibitory concentration against hrv-14 and the x-ray conformation of the compounds, a model has been developed which distinguishes between the space-filling properties of the active ... | 1990 | 2158559 |
| site-directed mutagenesis suggests close functional relationship between a human rhinovirus 3c cysteine protease and cellular trypsin-like serine proteases. | human rhinoviruses, like other picornaviruses, encode a cysteine protease (designated 3c) which cleaves mainly at viral gln-gly pairs. there are significant areas of homology between picornavirus 3c cysteine proteases and cellular serine proteases (e.g. trypsin), suggesting a functional relationship between their catalytic regions. to test this functional relationship, we made single substitutions in human rhinovirus type 14 protease 3c at seven amino acid positions which are highly conserved in ... | 1990 | 2158990 |
| comparative studies on the antirhinovirus activity and the mode of action of the rhinovirus capsid binding agents, chalcone amides. | studies of various analogs related to the antirhinovirus agent 4'-ethoxy-2'-hydroxy-4,6'-dimethoxychalcone (chalcone ro 09-0410) led to the identification of amide analogs that are 4.5 to 10 times more active against human rhinovirus (hrv) in tissue culture as measured by chemotherapeutic indices. chalcone amides ro 09-0535, ro 09-0696 and ro 09-0881 inhibited viral replication at concentrations as low as less than 2-3 ng/ml and were cytotoxic between 30 to 50 micrograms/ml. these compounds bind ... | 1990 | 2160796 |
| substrate requirements of human rhinovirus 3c protease for peptide cleavage in vitro. | a series of synthetic peptides representing authentic proteolytic cleavage sites of human rhinovirus type 14 were assayed as substrates for purified 3c protease. competition cleavage assays were employed to determine the relative specificity constants (kcat/km) for substrates with sequences related to the viral 2c-3a cleavage site. variable length peptides representing the 2c-3a cleavage site were cleaved with comparable efficiency. these studies defined a minimum substrate of 6 amino acids (tlf ... | 1990 | 2160953 |
| gross rearrangements within the 5'-untranslated region of the picornaviral genomes. | an analysis of reported nucleotide sequences revealed several cases of gross rearrangements in the 5'-untranslated region (5-utr) of picornaviral genomes. a large (greater than 100 nt) duplication was discovered in a downstream region of poliovirus 5-utr involved in the translational control. properties of the poliovirus mutants with large deletions [kuge and nomoto (1987) j. virol. 61, 1478-1487] show that a single copy of the appropriate repeating unit is compatible with a wild type phenotype ... | 1990 | 2162521 |
| pathogenicity for humans of human rhinovirus type 2 mutants resistant to or dependent on chalcone ro 09-0410. | mutants of human rhinovirus type 2 (hrv-2) resistant to and dependent on the antirhinoviral compound chalcone ro 09-0410 were selected in cell culture under clean laboratory conditions. a total of 42 volunteers were challenged with either the drug-resistant mutant [sr2-410(r)] (15 volunteers), the drug-dependent mutant [sr2-410(d)] (15 volunteers), or a wild-type hrv-2 which had a similar passage level in vitro as the mutants but without the drug (12 volunteers). of volunteers challenged with th ... | 1990 | 2168152 |
| crystallization and preliminary x-ray diffraction studies of the fab fragment of a neutralizing monoclonal antibody directed against human rhinovirus serotype 2. | we report on the preparation, crystallization, and preliminary x-ray diffraction analysis of the fab fragment of the monoclonal antibody 8f5 that neutralizes infectivity of human rhinovirus serotype 2 (hrv2). fab fragments prepared from this antibody by papain digestion were purified to isoelectric homogeneity by ion exchange chromatography and chromatofocusing. crystals were obtained by the hanging drop vapor diffusion method using ammonium sulfate as precipitant. the crystals belong to the ort ... | 1990 | 2170356 |
| inhibition of the uncoating of bovine enterovirus by short chain fatty acids. | short chain fatty acids inhibit the replication of bovine enterovirus but are almost ineffective against poliovirus type 1, coxsackievirus b5, encephalomyocarditis virus and human rhinovirus 1b. lauric acid binds to bovine enterovirus, thereby stabilizing the virus particle to heat degradation. fatty acid-bound virions attach to susceptible cells but fail to undergo cell-mediated uncoating. the inhibitory effect is reversible with chloroform and may result from a hydrophobic interaction between ... | 1990 | 2172446 |
| use of monoclonal antibodies to identify four neutralization immunogens on a common cold picornavirus, human rhinovirus 14. | a collection of 35 mouse monoclonal antibodies, raised against human rhinovirus 14 (hrv-14), was used to isolate 62 neutralization-resistant mutants. when cross-tested against the antibodies in a neutralization assay, the mutants fell into four antigenic groups, here called neutralization immunogens: nim-ia, -ib, -ii, and -iii. sequencing the mutant rna in segments corresponding to serotype-variable regions revealed that the amino acid substitutions segregated into clusters, which correlated exa ... | 1986 | 2416951 |
| a neutralizing epitope on human rhinovirus type 2 includes amino acid residues between 153 and 164 of virus capsid protein vp2. | use has been made of a monoclonal antibody (designated 8f5) to map a neutralizing epitope on the viral capsid protein vp2 of human rhinovirus 2 (hrv2). this antibody which was raised against the native virus, neutralizes hrv2 and is also capable of recognizing denatured vp2 on western blots. to examine the binding site of 8f5, vp2 of hrv2 was expressed in escherichia coli. deletions starting at the 3' end were then introduced into the gene for vp2 using bal-31 nuclease. polypeptides shortened at ... | 1987 | 2434607 |
| different rhinovirus serotypes neutralized by antipeptide antibodies. | recently, rossman et al. have described the three-dimensional structure of a human rhinovirus. a possible host cell surface receptor binding site was identified with a cleft on each icosahedral face. two highly conserved amino-acid sequences found in rhino-, polio-, and foot-and-mouth disease (fmd) viruses are located near the base of this site and could be important in maintaining its topology. we have prepared site-specific antibodies to two synthetic peptides which include these sequences. th ... | 1987 | 2444889 |
| use of crna probes for the detection of enteroviruses by molecular hybridization. | subgenomic fragments of cdna from poliovirus type 1 were inserted downstream from the sp6 or the t7 promoter in a gemini riboprobe vector and their in vitro synthesized rna transcripts were used as radiolabeled probes for the detection of enteroviral rnas by molecular hybridization. the crna transcripts appeared to be more sensitive probes than the corresponding cdnas. in vitro transcripts of the 5' noncoding region (5' nc riboprobe) were able to detect all of 14 reference enterovirus strains te ... | 1988 | 2448419 |
| antigenic sites on foot-and-mouth disease virus type a10. | a set of monoclonal antibodies was used to isolate nonneutralizable foot-and-mouth disease virus variants, and the rnas of the variants were sequenced. cross-neutralization studies and mapping of the amino acid changes indicated two major antigenic sites. the first site was trypsin sensitive and included the vp1 140 to 160 sequence. the second site was trypsin insensitive and included mainly vp3 residues. two minor sites were located near vp1 169 and on the c terminus of vp1. comparison with pol ... | 1988 | 2455819 |
| identification of an immunodominant antigenic site involving the capsid protein vp3 of hepatitis a virus. | hepatitis a virus, an hepatotropic picornavirus, is a common cause of acute hepatitis in man for which there is no available vaccine. competitive binding studies carried out in solid phase suggest that neutralizing monoclonal antibodies to hepatitis a virus recognize a limited number of epitopes on the capsid surface, although the polypeptide locations of these epitopes are not well defined. neutralization-escape mutants, selected for resistance to monoclonal antibodies, demonstrate broad cross- ... | 1988 | 2460866 |
| n-agib of poliovirus type 1: a discontinuous epitope formed by two loops of vp1 comprising residues 96-104 and 141-152. | analysis of resistant mutants to neutralizing monoclonal antibodies revealed a discontinuous neutralization epitope on vp1 of poliovirus type 1, mahoney. the epitope has the unique property of being also part of a sequential epitope within neutralization antigenic site i (n-agi). it is formed by residues in the loop 96-104 connecting the b and c strand and in the loop 141-152 connecting the d and e strand of vp1. because of strong analogy to neutralization immunogen ib (nimib) of human rhinoviru ... | 1989 | 2471354 |
| prediction of secondary structure, spatial organization and distribution of antigenic determinants for hepatitis a virus proteins. | on the basis of the secondary structure calculations from the known amino acid sequence we came to the conclusion that hepatitis a virus capsid proteins have the typical antiparallel beta-sheet bilayer structure. the predicted secondary structure of the hav proteins can be well aligned with those of the poliovirus (type 1 mahoney) and human rhinovirus (type 14). it enabled us to use the x-ray structure of the pv-1m and hrv-14 proteins as a template and then, firstly, to localize the positions of ... | 1987 | 2482756 |
| inhibition of rhinovirus attachment by neutralizing monoclonal antibodies and their fab fragments. | previous molecular and immunological studies have mapped four neutralization sites on human rhinovirus type 14 (b. sherry, a. g. mosser, r. j. colonno, and r. r. rueckert, j. virol. 57:246-257, 1986). eight monoclonal antibodies, one pair for each of the four target sites and all belonging to a single isotype, immunoglobulin g2a, were studied under conditions which resulted in 95% neutralization of infectious viral particles. all eight antibodies shifted the isoelectric point of virions from 6.7 ... | 1989 | 2535737 |
| human rhinovirus serotype 2: in vitro synthesis of an infectious rna. | a complete cdna copy of human rhinovirus serotype 2 rna was placed under the control of a t7 rna polymerase promoter. an in vitro transcribed rna containing two extra g residues at the 5' end gave rise to plaques on transfection into hela cells. the efficiency was approximately half that obtained with viral rna. on the contrary, an in vitro synthesized rna containing 16 additional nucleotides at the 5' end was not infectious. this ability to make an infectious in vitro transcribed rna will be us ... | 1989 | 2535899 |
| polypeptide 2a of human rhinovirus type 2: identification as a protease and characterization by mutational analysis. | evidence is presented that the protein 2a of human rhinovirus serotype 2 (hrv2) is a protease. on expression of the vp1-2a region of hrv2 in bacteria, protein 2a was capable of acting on its own n-terminus; derived extracts specifically cleaved a 16 amino acid oligopeptide corresponding to the sequence at the cleavage site. cleavage of the oligopeptide substrate provides a convenient in vitro assay system. deletion experiments showed that removal of 10 amino acids from the carboxy terminus inact ... | 1989 | 2538037 |
| the major human rhinovirus receptor is icam-1. | the major human rhinovirus receptor has been identified with monoclonal antibodies that inhibit rhinovirus infection. these monoclonal antibodies recognize a 95 kd cell surface glycoprotein on human cells and on mouse transfectants expressing a rhinovirus binding phenotype. purified 95 kd protein binds to rhinovirus in vitro. protein sequence from the 95 kd protein showed an identity with that of intercellular adhesion molecule-1 (icam-1); a cdna clone obtained from mouse transfectants expressin ... | 1989 | 2538243 |
| conformational change in the floor of the human rhinovirus canyon blocks adsorption to hela cell receptors. | a series of eight antiviral compounds complexed with human rhinovirus 14 (hrv-14) were previously shown to displace segments of polypeptide chains in the floor of the "canyon" by as much as 0.45 nm in c-alpha positions from the native conformation (j. badger, i. minor, m. j. kremer, m. a. oliveira, t. j. smith, j. p. griffith, d. m. a. guerin, s. krishnaswamy, m. luo, m. g. rossman, m. a. mckinlay, g. d. diana, f. j. dutko, m. fancher, r. r. rueckert, and b. a. heinz, proc. natl. acad. sci. usa ... | 1989 | 2539499 |
| evidence for the role of the p2 protein of human rhinovirus in its host range change. | human rhinovirus 39 (hrv39) is blocked in nonpermissive l cells at both adsorption and intracellular replication steps. we have selected a host range variant of hrv39 capable of bypassing the intracellular replication block and found it to have altered nonstructural p2 proteins. these alterations are similar to those reported earlier for host range variants of hrv2 (f. h. yin and n. b. lomax, j. virol. 48:410-418, 1983). this observation suggests that the intracellular replication block for both ... | 1989 | 2539529 |
| detection of serotypic variants in viral preparations: sensitivity of a procedure for selection. | the efficiency of an in vitro method (the breakthrough neutralization procedure) for selecting serotypic variants from preparations of human rhinovirus (hrv) 17 and a temperature sensitive strain (ts-1) of hrv-2 was examined. viruses were plaqued in the presence of homologous polyclonal antisera, and plaques which escaped neutralization were isolated. for control purposes, isolates were obtained by plaquing in the absence of antisera. any clone that consistently (minimum of 2 tests) yielded a 4- ... | 1989 | 2541156 |
| computer simulation study of the binding of an antiviral agent to a sensitive and a resistant human rhinovirus. | molecular dynamics simulations have been used to study the free energy of binding of an antiviral agent to the human rhinovirus hrv-14 and to a mutant in which a valine residue in the antiviral binding pocket is replaced by leucine. the simulations predict that the antiviral should bind to the two viruses with similar affinity, in apparent disagreement with experimental results. possible origins of this discrepancy are outlined. of particular importance is the apparent need for methods to system ... | 1989 | 2541225 |
| genetic and molecular analyses of spontaneous mutants of human rhinovirus 14 that are resistant to an antiviral compound. | spontaneous mutants of human rhinovirus 14 resistant to win 52084, an antiviral compound that inhibits attachment to cells, were isolated by selecting plaques that developed when wild-type virus was plated in the presence of high (2 micrograms/ml) or low (0.1 to 0.4 micrograms/ml) concentrations of the compound. two classes of drug resistance were observed: a high-resistance (hr) class with a frequency of about 4 x 10(-5), and a low-resistance (lr) class with a 10- to 30-fold-higher frequency. t ... | 1989 | 2542566 |
| three-dimensional structures of drug-resistant mutants of human rhinovirus 14. | mutants of human rhinovirus 14 were isolated and characterized by searching for resistance to compounds that inhibit viral uncoating. the portions of the rna that code for amino acids that surround the antiviral compound binding site were sequenced. x-ray analysis of two of these mutants, 1188 val----leu and 1199 cys----tyr, shows that these were single-site substitutions which would sterically hinder drug binding. differences in the resistance of mutant viruses to various antiviral compounds ma ... | 1989 | 2544734 |
| cdna cloning reveals that the major group rhinovirus receptor on hela cells is intercellular adhesion molecule 1. | a 90-kda surface glycoprotein was previously isolated and shown to be required for infection by the "major" group of human rhinovirus (hrv) serotypes. in the present work, the amino acid sequence of the receptor protein was obtained from cnbr and tryptic peptides. using degenerate oligonucleotides predicted from the peptide sequences, we identified four cdna clones that encode a 3-kilobase mrna. the clones were ligated, subcloned in a simian virus 40 expression vector, and used to transfect rece ... | 1989 | 2544880 |
| chimeric picornavirus polyproteins demonstrate a common 3c proteinase substrate specificity. | cross-species proteolytic processing was demonstrated by the 3c proteinases of human rhinovirus 14 and coxsackievirus b3 on poliovirus-specific polypeptide precursors. chimeric picornavirus cdna genomes were constructed in a t7 transcription vector in which the poliovirus 3c coding region was substituted with the corresponding allele from human rhinovirus 14 or coxsackievirus b3. in vitro translation and processing of the polypeptides encoded by the chimeric genomes demonstrated that the proteol ... | 1989 | 2545915 |
| use of synthetic oligonucleotide probes to detect rhinovirus rna. | current methods of detecting a human rhinovirus (hrv) infection are either based on isolation of virus in appropriate susceptible cell lines, which is time-consuming and requires considerable expertise, or are dependent on knowing the serotype. the existence of over 100 immunologically distinct serotypes makes serotype specific assays, such as elisa, unsuitable for general diagnostic assays. in this study a general rhinovirus assay is described which utilises synthetic oligonucleotides as probes ... | 1989 | 2546516 |
| the expression and purification of human rhinovirus protease 3c. | human rhinovirus type 14 protease 3c was expressed as a soluble and active protein in escherichia coli. the protease was purified by a cationic-exchange step followed by gel filtration on a tsk 3000 column. the final yield of purified protease was in the range 0.5-1.0 mg/l culture grown to a550 = 1.0. sequence analysis revealed that greater than 90% of the n-terminal residues were methionine. the enzyme activity of the purified protease was measured by cleavage of a synthetic peptide representin ... | 1989 | 2546760 |
| trypsin sensitivity of several human rhinovirus serotypes in their low ph-induced conformation. | five serotypes of human rhinovirus (hrv) were examined for sensitivity to trypsin at physiological ph, hrv1a, hrv2, and hrv14 were found to be resistant whereas in serotypes hrv49 and hrv89 degradation of vp2 was observed. however, exposure to low ph followed by neutralization, a treatment which causes irreversible conformational changes in the capsid, led to rapid cleavage by trypsin of vp1 in hrv1a, hrv2, and hrv49 at defined sites followed by degradation of vp2. in the case of hrv2, the cleav ... | 1989 | 2548332 |
| sequences in the 5' non-coding region of human rhinovirus 14 rna that affect in vitro translation. | a subgenomic cdna clone from human rhinovirus 14 (hrv-14), comprising the 5' non-coding region and the first 1182 nucleotides of the coding sequence, has been inserted into a vector under the control of the t7 promoter, and rna was transcribed. deletions in the 5' non-coding sequence modulated viral polyprotein synthesis significantly in a reticulocyte lysate system. removal of the first 491 nucleotides had little effect, but deletion of a further 55 nucleotides (491 to 546) significantly increa ... | 1989 | 2552008 |
| a theoretical study of the acidification of the rhinovirus capsid. | electrostatic calculations for human rhinovirus 14 indicate that histidine-base residue pairs in the region of a beta-strand interaction between pentamers may be involved in a ph-induced process that leads to the release of viral rna. other picornavirus sequences are examined for these residue pairs, a subset of which is present in enteroviruses. foot and mouth disease virus possesses one of the residue pairs, and cardioviruses, which undergo a separate ph and halide ion-induced capsid dissociat ... | 1989 | 2555222 |
| hydrolysis of a series of synthetic peptide substrates by the human rhinovirus 14 3c proteinase, cloned and expressed in escherichia coli. | the 3c proteins of several picornaviruses, including poliovirus, foot-and-mouth disease virus (fmdv) and encephalomyocarditis virus (emcv), have been demonstrated to be cysteine-type proteinases, involved in the processing of the respective polyproteins expressed by the monocistronic rna genome. nucleotide sequencing data have indicated that the human rhinovirus 14 (hrv-14) rna genome encodes a homologous 3c protein. the hrv-14 3c protein was purified to homogeneity from escherichia coli express ... | 1989 | 2555433 |
| typing of human rhinoviruses based on sequence variations in the 5' non-coding region. | unambiguous assignment of restriction enzyme patterns to six individual serotypes of human rhinovirus was accomplished after amplification of a 380 bp dna fragment derived from the 5' non-coding region. this was possible even though serotypes 1a and 1b and serotypes 2 and 49 differed only at 10 and 15 positions respectively. the method utilizes the conserved and variable components of this part of the genome and provides the basis for a simple and rapid method for typing of human rhinoviruses. | 1989 | 2555441 |
| selective elimination of interactions: a method for assessing thermodynamic contributions to ligand binding with application to rhinovirus antivirals. | a new method for evaluating the free energy of various physical interactions, such as hydrogen-bond, electrostatic, or van der waals interactions, is presented. rather than destroying or creating whole groups, selective (pairwise) interactions are eliminated from the total potential energy and the energy difference with the fully interacting system is evaluated. the exponential ensemble average of such an energy difference is then directly related to the corresponding free energy difference. thi ... | 1989 | 2555511 |
| crystal structure of human rhinovirus serotype 1a (hrv1a). | the structure of human rhinovirus 1a (hrv1a) has been determined to 3.2 a resolution using phase refinement and extension by symmetry averaging starting with phases at 5 a resolution calculated from the known human rhinovirus 14 (hrv14) structure. the polypeptide backbone structures of hrv1a and hrv14 are similar, but the exposed surfaces are rather different. differential charge distribution of amino acid residues in the "canyon", the putative receptor binding site, provides a possible explanat ... | 1989 | 2555523 |
| cleavage of small peptides in vitro by human rhinovirus 14 3c protease expressed in escherichia coli. | the 3c region of human rhinovirus 14 was expressed in escherichia coli. the microbially synthesized protease was functional, since the expressed precursor underwent autoproteolytic processing to generate mature molecules of the expected molecular weight and antigenicity. mutation of the putative active-site cys-146 residue to an alanine resulted in the synthesis of unprocessed precursor molecules. large quantities of the 20-kilodalton protease were purified by a simple purification protocol, and ... | 1989 | 2555540 |