| genome-wide analysis of protein-protein interactions and involvement of viral proteins in sars-cov replication. | analyses of viral protein-protein interactions are an important step to understand viral protein functions and their underlying molecular mechanisms. in this study, we adopted a mammalian two-hybrid system to screen the genome-wide intraviral protein-protein interactions of sars coronavirus (sars-cov) and therefrom revealed a number of novel interactions which could be partly confirmed by in vitro biochemical assays. three pairs of the interactions identified were detected in both directions: no ... | 2008 | 18827877 |
| t cell responses to whole sars coronavirus in humans. | effective vaccines should confer long-term protection against future outbreaks of severe acute respiratory syndrome (sars) caused by a novel zoonotic coronavirus (sars-cov) with unknown animal reservoirs. we conducted a cohort study examining multiple parameters of immune responses to sars-cov infection, aiming to identify the immune correlates of protection. we used a matrix of overlapping peptides spanning whole sars-cov proteome to determine t cell responses from 128 sars convalescent samples ... | 2008 | 18832706 |
| topology and membrane anchoring of the coronavirus replication complex: not all hydrophobic domains of nsp3 and nsp6 are membrane spanning. | coronaviruses express two very large replicase polyproteins, the 16 autoproteolytic cleavage products of which collectively form the membrane-anchored replication complexes. how these structures are assembled is still largely unknown, but it is likely that the membrane-spanning members of these nonstructural proteins (nsps) are responsible for the induction of the double-membrane vesicles and for anchoring the replication complexes to these membranes. for 3 of the 16 coronavirus nsps-nsp3, nsp4, ... | 2008 | 18842706 |
| evaluation of peptide-aldehyde inhibitors using r188i mutant of sars 3cl protease as a proteolysis-resistant mutant. | the 3c-like (3cl) protease of the severe acute respiratory syndrome (sars) coronavirus is a key enzyme for the virus maturation. we found for the first time that the mature sars 3cl protease is subject to degradation at 188arg/189gln. replacing arg with ile at position 188 rendered the protease resistant to proteolysis. the r188i mutant digested a conserved undecapeptide substrate with a k(m) of 33.8 microm and k(cat) of 4753 s(-1). compared with the value reported for the mature protease contai ... | 2008 | 18845442 |
| sars coronavirus: unusual lability of the nucleocapsid protein. | the severe acute respiratory syndrome (sars) is a contagious disease that killed hundreds and sickened thousands of people worldwide between november 2002 and july 2003. the nucleocapsid (n) protein of the coronavirus responsible for this disease plays a critical role in viral assembly and maturation and is of particular interest because of its potential as an antiviral target or vaccine candidate. refolding of sars n-protein during production and purification showed the presence of two addition ... | 2008 | 18926799 |
| interaction of severe acute respiratory syndrome-coronavirus and nl63 coronavirus spike proteins with angiotensin converting enzyme-2. | although in different groups, the coronaviruses severe acute respiratory syndrome-coronavirus (sars-cov) and nl63 use the same receptor, angiotensin converting enzyme (ace)-2, for entry into the host cell. despite this common receptor, the consequence of entry is very different; severe respiratory distress in the case of sars-cov but frequently only a mild respiratory infection for nl63. using a wholly recombinant system, we have investigated the ability of each virus receptor-binding protein, s ... | 2008 | 18931070 |
| [sars vaccines]. | severe acute respiratory syndrome (sars) is an emerging disease derived from wild animals and is highly pathogenic with a high mortality. a novel coronavirus sars coronavirus was identified to be a causative agent for sars. since its discovery, many trials have been executed to establish the prophylactic and therapeutics toward sars using laboratory animals. a number of different types of vaccines, such as inactivated virus vaccine, subunit vaccine, dna vaccine and vaccine using viral expression ... | 2008 | 18939499 |
| prior immunization with severe acute respiratory syndrome (sars)-associated coronavirus (sars-cov) nucleocapsid protein causes severe pneumonia in mice infected with sars-cov. | the details of the mechanism by which severe acute respiratory syndrome-associated coronavirus (sars-cov) causes severe pneumonia are unclear. we investigated the immune responses and pathologies of sars-cov-infected balb/c mice that were immunized intradermally with recombinant vaccinia virus (vv) that expressed either the sars-cov spike (s) protein (lc16m8rvv-s) or simultaneously all the structural proteins, including the nucleocapsid (n), membrane (m), envelope (e), and s proteins (lc16m8rvv- ... | 2008 | 18941225 |
| emerging and zoonotic infections in women. | emerging infections, many zoonotic, are caused by a variety of pathogens with global distribution. previously rare pathogens have emerged; global travel facilitates their rapid spread. human encroachment on remote areas has brought contact with zoonotic diseases never before characterized. although systematic study of rare outbreaks can be challenging, knowledge of emerging pathogens and their effects on women is accumulating. this article discusses effects of lymphocytic choriomeningitis virus, ... | 2008 | 18954762 |
| detection of antibodies against sars-coronavirus using recombinant truncated nucleocapsid proteins by elisa. | severe acute respiratory syndrome (sars) is a lifethreatening emerging respiratory disease caused by the coronavirus, sars-cov. the nucleocapsid (n) protein of sars-cov is highly antigenic and may be a suitable candidate for diagnostic applications. we constructed truncated recombinant n proteins (n1 [1-422 aa], n2 [1- 109 aa], and n3 [110-422 aa]) and determined their antigenicity by western blotting using convalescent sars serum. the recombinants containing n1 and n3 reacted with convalescent ... | 2008 | 18955825 |
| plp2, a potent deubiquitinase from murine hepatitis virus, strongly inhibits cellular type i interferon production. | infections by coronaviruses such as severe acute respiratory syndrome (sars) coronavirus (scov) and mouse hepatitis virus a59 (mhv-a59) result in very little type i interferon (ifn) production by host cells, which is potentially responsible for the rapid viral growth and severe immunopathology associated with sars. however, the molecular mechanisms for the low ifn production in cells infected with coronaviruses remain unclear. here, we provide evidence that papain-like protease domain 2 (plp2), ... | 2008 | 18957937 |
| expression of sars-coronavirus spike glycoprotein in pichia pastoris. | to establish a rapid and economical method for the expression of viral proteins in high yield and purity by pichia pastoris, the s protein of the sars-cov was selected in this study. six s glycoprotein fragments were expressed in escherichia coli bl21 and yeast km71h strains. after purification by affinity chromatography, the protein identities were confirmed by western blot analysis, n-terminal sequencing and mass spectrometry. the proteins expressed in e. coli were low in solubility and bound ... | 2009 | 18958613 |
| protease-mediated entry via the endosome of human coronavirus 229e. | human coronavirus 229e, classified as a group i coronavirus, utilizes human aminopeptidase n (apn) as a receptor; however, its entry mechanism has not yet been fully elucidated. we found that hela cells infected with 229e via apn formed syncytia when treated with trypsin or other proteases but not in a low-ph environment, a finding consistent with syncytium formation by severe acute respiratory syndrome coronavirus (sars-cov). in addition, trypsin induced cleavage of the 229e s protein. by using ... | 2009 | 18971274 |
| differential activities of cellular and viral macro domain proteins in binding of adp-ribose metabolites. | macro domain is a highly conserved protein domain found in both eukaryotes and prokaryotes. macro domains are also encoded by a set of positive-strand rna viruses that replicate in the cytoplasm of animal cells, including coronaviruses and alphaviruses. the functions of the macro domain are poorly understood, but it has been suggested to be an adp-ribose-binding module. we have here characterized three novel human macro domain proteins that were found to reside either in the cytoplasm and nucleu ... | 2009 | 18983849 |
| structural and biochemical investigation of heptad repeat derived peptides of human sars corona virus (hsars-cov) spike protein. | hsars-cov is the causative agent for sars infection. its spike glycoprotein (s) is processed by host furin enzyme to produce s1 and s2 fragments, the latter being crucial for fusion with the host membrane. this takes place via formation of a coiled coil 6-helix bundle involving n and c-terminal heptad repeat domains (hr-n and hr-c) of s2. several fluorescent and non-fluorescent peptides from these domains were synthesized to examine their interactions by circular dichroism, thermal denaturation, ... | 2008 | 18991761 |
| detection and phylogenetic analysis of group 1 coronaviruses in south american bats. | bat coronaviruses (bt-covs) are thought to be the precursors of severe acute respiratory syndrome coronavirus. we detected bt-covs in 2 bat species from trinidad. phylogenetic analysis of the rna-dependent rna polymerase gene and helicase confirmed them as group 1 coronaviruses. | 2008 | 19046513 |
| nuclear magnetic resonance structure shows that the severe acute respiratory syndrome coronavirus-unique domain contains a macrodomain fold. | the nuclear magnetic resonance (nmr) structure of a central segment of the previously annotated severe acute respiratory syndrome (sars)-unique domain (sud-m, for "middle of the sars-unique domain") in sars coronavirus (sars-cov) nonstructural protein 3 (nsp3) has been determined. sud-m(513-651) exhibits a macrodomain fold containing the nsp3 residues 528 to 648, and there is a flexibly extended n-terminal tail with the residues 513 to 527 and a c-terminal flexible tail of residues 649 to 651. a ... | 2009 | 19052085 |
| novel influenza virus ns1 antagonists block replication and restore innate immune function. | the innate immune system guards against virus infection through a variety of mechanisms including mobilization of the host interferon system, which attacks viral products mainly at a posttranscriptional level. the influenza virus ns1 protein is a multifunctional facilitator of virus replication, one of whose actions is to antagonize the interferon response. since ns1 is required for efficient virus replication, it was reasoned that chemical inhibitors of this protein could be used to further und ... | 2009 | 19052087 |
| mechanisms of severe acute respiratory syndrome pathogenesis and innate immunomodulation. | the modulation of the immune response is a common practice of many highly pathogenic viruses. the emergence of the highly pathogenic coronavirus severe acute respiratory virus (sars-cov) serves as a robust model system to elucidate the virus-host interactions that mediate severe end-stage lung disease in humans and animals. coronaviruses encode the largest positive-sense rna genome of approximately 30 kb, encode a variety of replicase and accessory open reading frames that are structurally uniqu ... | 2008 | 19052324 |
| characteristic features and outcomes of severe acute respiratory syndrome found in severe acute respiratory syndrome intensive care unit patients. | the aim of the study was to identify characteristic clinical features and outcomes of critically ill patients with confirmed severe acute respiratory syndrome (sars). | 2008 | 19056023 |
| detection of sars coronavirus in humans and animals by conventional and quantitative (real time) reverse transcription polymerase chain reactions. | severe acute respiratory syndrome is a novel human disease caused by a coronavirus of animal origin. soon after the discovery sars-cov, several molecular assays were described for the detection of this virus. of these, conventional and quantitative rt-pcr approaches were the primary tools for sars-cov rna detection. in this chapter we describe a two-step conventional rt-pcr and a one-step quantitative rt-pcr that were used routinely in our laboratories during the sars outbreak. | 2008 | 19057863 |
| pseudotyped vesicular stomatitis virus for analysis of virus entry mediated by sars coronavirus spike proteins. | severe acute respiratory syndrome (sars) coronavirus (cov) contains a spike (s) protein that binds to a receptor molecule (angiotensin-converting enzyme 2; ace2), induces membrane fusion, and serves as a neutralizing epitope. to study the functions of the s protein, we describe here the generation of sars-cov s protein-bearing vesicular stomatitis virus (vsv) pseudotype using a vsvdeltag*/gfp system in which the g gene is replaced by the green fluorescent protein (gfp) gene (vsv-sars-cov-st19/gf ... | 2008 | 19057867 |
| engineering infectious cdnas of coronavirus as bacterial artificial chromosomes. | the construction of coronavirus (cov) infectious clones had been hampered by the large size of the viral genome (around 30kb) and the instability of plasmids carrying cov replicase sequences in escherichia coli. several approaches have been developed to overcome these problems. here we describe the engineering of cov full-length cdna clones using bacterial artificial chromosomes (bacs). in this system the viral rna is expressed in the cell nucleus under the control of the cytomegalovirus promote ... | 2008 | 19057870 |
| detection of group 1 coronaviruses in bats using universal coronavirus reverse transcription polymerase chain reactions. | the zoonotic transmission of sars coronavirus from animals to humans revealed the potential impact of coronaviruses on mankind. this incident also triggered several surveillance programs to hunt for novel coronaviruses in human and wildlife populations. using classical rt-pcr assays that target a highly conserved sequence among coronaviruses, we identified the first coronaviruses in bats. these assays and the cloning and sequencing of the pcr products are described in this chapter. using the sam ... | 2008 | 19057871 |
| generation of recombinant coronaviruses using vaccinia virus as the cloning vector and stable cell lines containing coronaviral replicon rnas. | coronavirus reverse genetic systems have become valuable tools for studying the molecular biology of coronavirus infections. they have been applied to the generation of recombinant coronaviruses, selectable replicon rnas, and coronavirus-based vectors for heterologous gene expression. here we provide a collection of protocols for the generation, cloning, and modification of full-length coronavirus cdna using vaccinia virus as a cloning vector. based on cloned coronaviral cdna, we describe the ge ... | 2008 | 19057873 |
| establishment and characterization of monoclonal antibodies against sars coronavirus. | immunological detection of viruses and their components by monoclonal antibodies is a powerful method for studying the structure and function of viral molecules. here we describe detailed methods for establishing monoclonal antibodies against severe acute respiratory syndrome coronavirus (sars-cov). b cell hybridomas are generated from mice that are hyperimmunized with inactivated sars-cov virions. the hybridomas produce monoclonal antibodies that recognize viral component molecules, including t ... | 2008 | 19057876 |
| large-scale preparation of uv-inactivated sars coronavirus virions for vaccine antigen. | in general, a whole virion serves as a simple vaccine antigen and often essential material for the analysis of immune responses against virus infection. however, to work with highly contagious pathogens, it is necessary to take precautions against laboratory-acquired infection. we have learned many lessons from the recent outbreak of severe acute respiratory syndrome (sars). in order to develop an effective vaccine and diagnostic tools, we prepared uv-inactivated sars coronavirus on a large scal ... | 2008 | 19057880 |
| school model and new targeting strategies. | protein-protein interactions play a central role in biological processes and thus are an appealing target for innovative drug design a nd development. they can be targeted bysmall molecule inhibitors, peptides and peptidomimetics, which represent an alternative to protein therapeutics that carry many disadvantages. in this chapter, i describe specific protein-protein interactions suggested by a novel model of immune signaling, the signaling chain homooligomerization (school) model, to be critica ... | 2008 | 19065798 |
| sars- and other coronaviruses. | | 2008 | 19068525 |
| myd88 is required for protection from lethal infection with a mouse-adapted sars-cov. | a novel human coronavirus, sars-cov, emerged suddenly in 2003, causing approximately 8000 human cases and more than 700 deaths worldwide. since most animal models fail to faithfully recapitulate the clinical course of sars-cov in humans, the virus and host factors that mediate disease pathogenesis remain unclear. recently, our laboratory and others developed a recombinant mouse-adapted sars-cov (rma15) that was lethal in balb/c mice. in contrast, intranasal infection of young 10-week-old c57bl/6 ... | 2008 | 19079579 |
| globalisation and blood safety. | globalisation may be viewed as the growing interdependence of countries worldwide through the increasing volume and variety of cross-border transactions in goods and services, and also through the more rapid and widespread diffusion of technology. globalisation is not just an economic phenomenon, although it is frequently described as such, but includes commerce, disease and travel, and immigration, and as such it affects blood safety and supply in various ways. the relatively short travel times ... | 2009 | 19081166 |
| severe acute respiratory syndrome coronavirus protein 6 is required for optimal replication. | severe acute respiratory syndrome coronavirus (sars-cov) encodes several accessory proteins of unknown function. one of these proteins, protein 6 (p6), which is encoded by orf6, enhances virus replication when introduced into a heterologous murine coronavirus (mouse hepatitis virus [mhv]) but is not essential for optimal sars-cov replication after infection at a relatively high multiplicity of infection (moi). here, we reconcile these apparently conflicting results by showing that p6 enhances sa ... | 2009 | 19091867 |
| glycogen synthase kinase-3 regulates the phosphorylation of severe acute respiratory syndrome coronavirus nucleocapsid protein and viral replication. | coronavirus (cov) nucleocapsid (n) protein is a highly phosphorylated protein required for viral replication, but whether its phosphorylation and the related kinases are involved in the viral life cycle is unknown. we found the severe acute respiratory syndrome cov n protein to be an appropriate system to address this issue. using high resolution page analysis, this protein could be separated into phosphorylated and unphosphorylated isoforms. mass spectrometric analysis and deletion mapping show ... | 2009 | 19106108 |
| severe acute respiratory syndrome coronavirus triggers apoptosis via protein kinase r but is resistant to its antiviral activity. | in this study, infection of 293/ace2 cells with severe acute respiratory syndrome coronavirus (sars-cov) activated several apoptosis-associated events, namely, cleavage of caspase-3, caspase-8, and poly(adp-ribose) polymerase 1 (parp), and chromatin condensation and the phosphorylation and hence inactivation of the eukaryotic translation initiation factor 2alpha (eif2alpha). in addition, two of the three cellular eif2alpha kinases known to be virus induced, protein kinase r (pkr) and pkr-like en ... | 2009 | 19109397 |
| the pathogen receptor liver and lymph node sinusoidal endotelial cell c-type lectin is expressed in human kupffer cells and regulated by pu.1. | human lsectin (liver and lymph node sinusoidal endothelial cell c-type lectin, clec4g) is a c-type lectin encoded within the l-sign/dc-sign/cd23 gene cluster. lsectin acts as a pathogen attachment factor for ebolavirus and the sars coronavirus, and its expression can be induced by interleukin-4 on monocytes and macrophages. although reported as a liver and lymph node sinusoidal endothelial cell-specific molecule, lsectin could be detected in the mutz-3 dendritic-like cell line at the messenger r ... | 2009 | 19111020 |
| sars-cov proteins decrease levels and activity of human enac via activation of distinct pkc isoforms. | among the multiple organ disorders caused by the severe acute respiratory syndrome coronavirus (sars-cov), acute lung failure following atypical pneumonia is the most serious and often fatal event. we hypothesized that two of the hydrophilic structural coronoviral proteins (s and e) would regulate alveolar fluid clearance by decreasing the cell surface expression and activity of amiloride-sensitive epithelial sodium (na(+)) channels (enac), the rate-limiting protein in transepithelial na(+) vect ... | 2009 | 19112100 |
| severe acute respiratory syndrome-associated coronavirus infection in toronto children: a second look. | during the severe acute respiratory syndrome outbreak of 2003, there was an impetus to provide clinical information to the medical community in a timely manner. accordingly, a preliminary report of our experience of suspected severe acute respiratory syndrome-associated coronavirus infections in children was published without microbiological findings. this report provides an update on pediatric severe acute respiratory syndrome-associated coronavirus infections in toronto, ontario, canada, that ... | 2009 | 19117866 |
| analysis of severe acute respiratory syndrome coronavirus structural proteins in virus-like particle assembly. | sars-cov has four major structural proteins: the n, s, m, and e proteins. to investigate the mechanism of sars-cov assembly, we cloned the genes encoding these four proteins into the eukaryotic expression vector pcaggs and transfected them into 293t cells. when all four expression vectors were co-transfected vlp formed, as confirmed using electron microscopy. using a rabbit polyclonal antibody specific to the n protein, n-protein-containing particles similar in size to the vlp were also observed ... | 2008 | 19120977 |
| crossing barriers: infections of the lung and the gut. | although known as respiratory pathogens, severe acute respiratory syndrome (sars) and its sister coronaviruses frequently cause enteric symptoms. in addition, other classically non-enteric viruses (such as hiv and influenza) may also have enteric effects that are crucial in their pathogeneses. these effects can be due to direct infection of the gut mucosa, but can also be because of decreased antibacterial defenses, increased mucosal permeability, bacterial translocation, and systemic leak of en ... | 2009 | 19129753 |
| structural basis of inhibition specificities of 3c and 3c-like proteases by zinc-coordinating and peptidomimetic compounds. | human coxsackievirus (cv) belongs to the picornavirus family, which consists of over 200 medically relevant viruses. in picornavirus, a chymotrypsin-like protease (3c(pro)) is required for viral replication by processing the polyproteins, and thus it is regarded as an antiviral drug target. a 3c-like protease (3cl(pro)) also exists in human coronaviruses (cov) such as 229e and the one causing severe acute respiratory syndrome (sars). to combat sars, we previously had developed peptidomimetic and ... | 2009 | 19144641 |
| peptide nanoparticles as novel immunogens: design and analysis of a prototypic severe acute respiratory syndrome vaccine. | severe acute respiratory syndrome (sars) is an infectious disease caused by a novel coronavirus that cost nearly 800 lives. while there have been no recent outbreaks of the disease, the threat remains as sars coronavirus (sars-cov) like strains still exist in animal reservoirs. therefore, the development of a vaccine against sars is in grave need. here, we have designed and produced a prototypic sars vaccine: a self-assembling polypeptide nanoparticle that repetitively displays a sars b-cell epi ... | 2009 | 19152635 |
| severe acute respiratory syndrome coronavirus nsp9 dimerization is essential for efficient viral growth. | the severe acute respiratory syndrome coronavirus (sars-cov) devotes a significant portion of its genome to producing nonstructural proteins required for viral replication. sars-cov nonstructural protein 9 (nsp9) was identified as an essential protein with rna/dna-binding activity, and yet its biological function within the replication complex remains unknown. nsp9 forms a dimer through the interaction of parallel alpha-helices containing the protein-protein interaction motif gxxxg. in order to ... | 2009 | 19153232 |
| a novel replication-competent vaccinia vector mvtt is superior to mva for inducing high levels of neutralizing antibody via mucosal vaccination. | mucosal vaccination offers great advantage for inducing protective immune response to prevent viral transmission and dissemination. here, we report our findings of a head-to-head comparison of two viral vectors modified vaccinia ankara (mva) and a novel replication-competent modified vaccinia tian tan (mvtt) for inducing neutralizing antibodies (nabs) via intramuscular and mucosal vaccinations in mice. mvtt is an attenuated variant of the wild-type vtt, which was historically used as a smallpox ... | 2009 | 19159014 |
| a noncovalent class of papain-like protease/deubiquitinase inhibitors blocks sars virus replication. | we report the discovery and optimization of a potent inhibitor against the papain-like protease (plpro) from the coronavirus that causes severe acute respiratory syndrome (sars-cov). this unique protease is not only responsible for processing the viral polyprotein into its functional units but is also capable of cleaving ubiquitin and isg15 conjugates and plays a significant role in helping sars-cov evade the human immune system. we screened a structurally diverse library of 50,080 compounds for ... | 2008 | 18852458 |
| an efficient method for recovery of target ssdna based on amino-modified silica-coated magnetic nanoparticles. | in this paper, an improved recovery method for target ssdna using amino-modified silica-coated magnetic nanoparticles (asmnps) is reported. this method takes advantages of the amino-modified silica-coated magnetic nanoparticles prepared using water-in-oil microemulsion technique, which employs amino-modified silica as the shell and iron oxide as the core of the magnetic nanoparticles. the nanoparticles have a silica surface with amino groups and can be conjugated with any desired bio-molecules t ... | 2005 | 18970204 |
| seroprevalence of sars coronavirus among residents near a hospital with a nosocomial outbreak. | an epidemic of severe acute respiratory syndrome (sars) occurred in taiwan from april to july 2003. a nosocomial outbreak of sars occurred at kaohsiung chang gung memorial hospital (cgmh) in may 2003. the purpose of our study was to survey the prevalence of the sars coronavirus (cov) in a community adjacent to kaohsiung cgmh and collect demographic data, including basic information about health status, household, and possible risk factors for sars-cov infection. | 2008 | 18971158 |
| identification of a minimal peptide derived from heptad repeat (hr) 2 of spike protein of sars-cov and combination of hr1-derived peptides as fusion inhibitors. | the heptad repeats (hr1 and hr2) of the spike protein of sars-cov are highly conserved regions forming a critical 6-helix bundle during the fusion step of virus entry and are attractive targets of entry inhibitors. in this study, we report that a minimal hr2 peptide, p6 of 23-mer, can block the fusion of sars-cov with an ic(50) of 1.04+/-0.22 microm. this finding supports the structural prediction of the deep groove of hr1 trimer as a target for fusion inhibitors, and suggests p6 as a potential ... | 2009 | 18983873 |
| conserved amino acids w423 and n424 in receptor-binding domain of sars-cov are potential targets for therapeutic monoclonal antibody. | the receptor-binding domain (rbd) on spike protein of severe acute respiratory syndrome-associated coronavirus (sars-cov) is the main region interacting with the viral receptor-ace2 and is a useful target for induction of neutralizing antibodies against sars-cov infection. here we generated two monoclonal antibodies (mabs), targeting rbd, with marked virus neutralizing activity. the mabs recognize a new conformational epitope which consists of several discontinuous peptides (aa. 343-367, 373-390 ... | 2009 | 18986662 |
| crystal structures of two coronavirus adp-ribose-1''-monophosphatases and their complexes with adp-ribose: a systematic structural analysis of the viral adrp domain. | the coronaviruses are a large family of plus-strand rna viruses that cause a wide variety of diseases both in humans and in other organisms. the coronaviruses are composed of three main lineages and have a complex organization of nonstructural proteins (nsp's). in the coronavirus, nsp3 resides a domain with the macroh2a-like fold and adp-ribose-1"-monophosphatase (adrp) activity, which is proposed to play a regulatory role in the replication process. however, the significance of this domain for ... | 2009 | 18987156 |
| humoral and cellular immune responses induced by 3a dna vaccines against severe acute respiratory syndrome (sars) or sars-like coronavirus in mice. | vaccine development for severe acute respiratory syndrome coronavirus (sars-cov) has mainly focused on the spike (s) protein. however, the variation of the s gene between viruses may affect the efficacy of a vaccine, particularly for cross-protection against sars-like cov (sl-cov). recently, a more conserved group-specific open reading frame (orf), the 3a gene, was found in both sars-cov and sl-cov. here, we studied the immunogenicity of human sars-cov 3a and bat sl-cov 3a dna vaccines in mice t ... | 2009 | 18987164 |
| simple tests for rapid detection of canine parvovirus antigen and canine parvovirus-specific antibodies. | canine parvovirus (cpv) is the number one viral cause of enteritis, morbidity, and mortality in 8-week-old young puppies. we have developed twin assays (slide agglutination test [sat] for cpv antigen and slide inhibition test [sit] for cpv antibody) that are sensitive, specific, cost-effective, generic for all genotypes of cpv, and provide instant results for cpv antigen detection in feces and antibody quantification in serum. we found these assays to be useful for routine applications in kennel ... | 2009 | 18987166 |
| broadening of neutralization activity to directly block a dominant antibody-driven sars-coronavirus evolution pathway. | phylogenetic analyses have provided strong evidence that amino acid changes in spike (s) protein of animal and human sars coronaviruses (sars-covs) during and between two zoonotic transfers (2002/03 and 2003/04) are the result of positive selection. while several studies support that some amino acid changes between animal and human viruses are the result of inter-species adaptation, the role of neutralizing antibodies (nabs) in driving sars-cov evolution, particularly during intra-species transm ... | 2008 | 18989460 |
| severe acute respiratory syndrome (sars) coronavirus-induced lung epithelial cytokines exacerbate sars pathogenesis by modulating intrinsic functions of monocyte-derived macrophages and dendritic cells. | severe acute respiratory syndrome (sars), which is caused by a novel coronavirus (cov), is a highly communicable disease with the lungs as the major pathological target. although sars likely stems from overexuberant host inflammatory responses, the exact mechanism leading to the detrimental outcome in patients remains unknown. pulmonary macrophages (mphi), airway epithelium, and dendritic cells (dc) are key cellular elements of the host innate defenses against respiratory infections. while pulmo ... | 2009 | 19004938 |
| turkey coronavirus non-structure protein nsp15--an endoribonuclease. | turkey coronavirus (tcov) polyprotein was predicted to be cleaved into 15 non-structural proteins (nsp2 to nsp16), but none of these nsps have been characterized. tcov nsp15 consists of 338 residues and shares 40% sequence similarity to u-specific nidovirales endoribonuclease (nendou) of severe acute respiratory syndrome coronavirus. objective: the purpose of the present study was to characterize tcov nsp15. methods: the tcov nsp15 gene was cloned into ptriex1 and expressed as a c-terminal his-t ... | 2008 | 19023218 |
| differential inhibitory activities and stabilisation of dna aptamers against the sars coronavirus helicase. | the helicase from severe acute respiratory syndrome coronavirus (sars-cov) possesses ntpase, duplex rna/dna-unwinding and rna-capping activities that are essential for viral replication and proliferation. here, we have isolated dna aptamers against the sars-cov helicase from a combinatorial dna library. these aptamers show two distinct classes of secondary structure, g-quadruplex and non-g-quadruplex, as shown by circular dichroism and gel electrophoresis. all of the aptamers that were selected ... | 2008 | 19031435 |
| synthetic recombinant bat sars-like coronavirus is infectious in cultured cells and in mice. | defining prospective pathways by which zoonoses evolve and emerge as human pathogens is critical for anticipating and controlling both natural and deliberate pandemics. however, predicting tenable pathways of animal-to-human movement has been hindered by challenges in identifying reservoir species, cultivating zoonotic organisms in culture, and isolating full-length genomes for cloning and genetic studies. the ability to design and recover pathogens reconstituted from synthesized cdnas has the p ... | 2008 | 19036930 |
| the expression and antigenicity of a truncated spike-nucleocapsid fusion protein of severe acute respiratory syndrome-associated coronavirus. | in the absence of effective drugs, controlling sars relies on the rapid identification of cases and appropriate management of the close contacts, or effective vaccines for sars. therefore, developing specific and sensitive laboratory tests for sars as well as effective vaccines are necessary for national authorities. | 2008 | 19038059 |
| development of an enzyme-linked immunosorbent assay-based test with a cocktail of nucleocapsid and spike proteins for detection of severe acute respiratory syndrome-associated coronavirus-specific antibody. | a new enzyme-linked immunosorbent assay (elisa)-based immunoglobulin g (igg)-plus-igm antibody detection test for severe acute respiratory syndrome (sars) has been developed by using a cocktail of four recombinant polypeptides as the antigen. these recombinant fragments were designed as parts of two different structural proteins from sars-associated coronavirus (sars-cov). one recombinant polypeptide, s251-683, was designed as part of the spike glycoprotein, and the other three polypeptides comp ... | 2009 | 19038782 |
| viral protease inhibitors. | this review provides an overview of the development of viral protease inhibitors as antiviral drugs. we concentrate on hiv-1 protease inhibitors, as these have made the most significant advances in the recent past. thus, we discuss the biochemistry of hiv-1 protease, inhibitor development, clinical use of inhibitors, and evolution of resistance. since many different viruses encode essential proteases, it is possible to envision the development of a potent protease inhibitor for other viruses if ... | 2009 | 19048198 |
| searching immunodominant epitopes prior to epidemic: hla class ii-restricted sars-cov spike protein epitopes in unexposed individuals. | identification of dominant t cell epitopes within newly emerging and re-emerging infectious organisms is valuable in understanding pathogenic immune responses and potential vaccine designs. however, difficulties in obtaining samples from patients or convalescent subjects have hampered research in this direction. we demonstrated a strategy, tetramer-guided epitope mapping, that specific cd4+ t cell epitopes can be identified by using pbmc from subjects that have not been exposed to the infectious ... | 2009 | 19050106 |
| multiple nucleic acid binding sites and intrinsic disorder of severe acute respiratory syndrome coronavirus nucleocapsid protein: implications for ribonucleocapsid protein packaging. | the nucleocapsid protein (n) of the severe acute respiratory syndrome coronavirus (sars-cov) packages the viral genomic rna and is crucial for viability. however, the rna-binding mechanism is poorly understood. we have shown previously that the n protein contains two structural domains--the n-terminal domain (ntd; residues 45 to 181) and the c-terminal dimerization domain (ctd; residues 248 to 365)--flanked by long stretches of disordered regions accounting for almost half of the entire sequence ... | 2009 | 19052082 |
| the identification of a calmodulin-binding domain within the cytoplasmic tail of angiotensin-converting enzyme-2. | angiotensin-converting enzyme (ace)-2 is a homolog of the well-characterized plasma membrane-bound angiotensin-converting enzyme. ace2 is thought to play a critical role in regulating heart function, and in 2003, ace2 was identified as a functional receptor for severe acute respiratory syndrome coronavirus. we have recently shown that like ace, ace2 undergoes ectodomain shedding and that this shedding event is up-regulated by phorbol esters. in the present study, we used gel shift assays to demo ... | 2009 | 19164471 |
| the identification of a calmodulin-binding domain within the cytoplasmic tail of angiotensin-converting enzyme-2. | angiotensin-converting enzyme (ace)-2 is a homolog of the well-characterized plasma membrane-bound angiotensin-converting enzyme. ace2 is thought to play a critical role in regulating heart function, and in 2003, ace2 was identified as a functional receptor for severe acute respiratory syndrome coronavirus. we have recently shown that like ace, ace2 undergoes ectodomain shedding and that this shedding event is up-regulated by phorbol esters. in the present study, we used gel shift assays to demo ... | 2009 | 19164471 |
| delivery to the lower respiratory tract is required for effective immunization with newcastle disease virus-vectored vaccines intended for humans. | newcastle disease virus (ndv), an avian virus, is being evaluated for the development of vectored human vaccines against emerging pathogens. previous studies of ndv-vectored vaccines in a mouse model suggested their potency after delivery by injection or by the intranasal route. we compared the efficacy of various routes of delivery of ndv-vectored vaccines in a non-human primate model. while delivery of an ndv-vectored vaccine by the combined intranasal/intratracheal route elicited protective i ... | 2009 | 19168110 |
| crystal structures of the x-domains of a group-1 and a group-3 coronavirus reveal that adp-ribose-binding may not be a conserved property. | the polyproteins of coronaviruses are cleaved by viral proteases into at least 15 nonstructural proteins (nsps). consisting of five domains, nsp3 is the largest of these (180-210 kda). among these domains, the so-called x-domain is believed to act as adp-ribose-1''-phosphate phosphatase or to bind poly(adp-ribose). however, here we show that the x-domain of infectious bronchitis virus (strain beaudette), a group-3 coronavirus, fails to bind adp-ribose. this is explained on the basis of the cryst ... | 2009 | 19177346 |
| sars coronavirus spike protein-induced innate immune response occurs via activation of the nf-kappab pathway in human monocyte macrophages in vitro. | a purified recombinant spike (s) protein was studied for its effect on stimulating human peripheral blood monocyte macrophages (pbmc). we examined inflammatory gene mrna abundances found in s protein-treated pbmc using gene arrays. we identified differential mrna abundances of genes with functional properties associated with antiviral (cxcl10) and inflammatory (il-6 and il-8) responses. we confirmed cytokine mrna increases by real-time quantitative(q) rt-pcr or elisa. we further analyzed the sen ... | 2009 | 19185596 |
| comparative analysis of the immunogenicity of sars-cov nucleocapsid dna vaccine administrated with different routes in mouse model. | the development of strategies to augment the immunogenicity of dna vaccines is critical for improving their clinical utility. one such strategy involves using the different immune routes with dna vaccines. in the present study, the immunogenicity of sars-cov nucleocapsid dna vaccine, induced by using the current routine vaccination routes (intramuscularly, by electroporation, or orally using live-attenuated salmonella typhimurium), was compared in mouse model. the comparison between the three va ... | 2009 | 19186202 |
| the spike protein of sars-cov--a target for vaccine and therapeutic development. | severe acute respiratory syndrome (sars) is a newly emerging infectious disease caused by a novel coronavirus, sars-coronavirus (sars-cov). the sars-cov spike (s) protein is composed of two subunits; the s1 subunit contains a receptor-binding domain that engages with the host cell receptor angiotensin-converting enzyme 2 and the s2 subunit mediates fusion between the viral and host cell membranes. the s protein plays key parts in the induction of neutralizing-antibody and t-cell responses, as we ... | 2009 | 19198616 |
| molecular targets for diagnostics and therapeutics of severe acute respiratory syndrome (sars-cov). | the large number of deaths in a short period of time due to the spread of severe acute respiratory syndrome (sars) infection led to the unparalleled collaborative efforts world wide to determine and characterize the new coronavirus (sars-cov). the full genome sequence was determined within weeks of the first outbreak by the canadian group with international collaboration. as per the world health organization (who), the continual lack of a rapid laboratory test to aid the early diagnosis of suspe ... | 2008 | 19203466 |
| a computational analysis of sars cysteine proteinase-octapeptide substrate interaction: implication for structure and active site binding mechanism. | sars coronavirus main proteinase (sars covmpro) is an important enzyme for the replication of severe acute respiratory syndrome virus. the active site region of sars covmpro is divided into 8 subsites. understanding the binding mode of sars covmpro with a specific substrate is useful and contributes to structural-based drug design. the purpose of this research is to investigate the binding mode between the sars covmpro and two octapeptides, especially in the region of the s3 subsite, through a m ... | 2009 | 19208150 |
| functional screen reveals sars coronavirus nonstructural protein nsp14 as a novel cap n7 methyltransferase. | the n7-methylguanosine (m7g) cap is the defining structural feature of eukaryotic mrnas. most eukaryotic viruses that replicate in the cytoplasm, including coronaviruses, have evolved strategies to cap their rnas. in this report, we used a yeast genetic system to functionally screen for the cap-forming enzymes encoded by severe acute respiratory syndrome (sars) coronavirus and identified the nonstructural protein (nsp) 14 of sars coronavirus as a (guanine-n7)-methyltransferase (n7-mtase) in vivo ... | 2009 | 19208801 |
| sars-cov spike proteins expressed by the vaccinia virus tiantan strain: secreted sq protein induces robust neutralization antibody in mice. | the spike (s) glycoprotein of severe acute respiratory syndrome coronavirus (sars-cov) is a major target in the development of diagnostic assays and vaccines, but its antigenic and immunogenic properties remain unclear. seven sars-cov spike proteins (s, sq, s1, rbd, s2, s2q, and cx) were generated using the modified vaccinia virus (tiantan strain) as a vector, and their antigenicity and immunogenicity were evaluated. the secreted sq protein in which the transmembrane domain was deleted, as well ... | 2009 | 19210229 |
| potent human monoclonal antibodies against sars cov, nipah and hendra viruses. | background: recently, several potently neutralizing fully human monoclonal antibodies (hmabs) targeting the severe acute respiratory syndrome-associated coronavirus (sars cov) s glycoprotein, and the g glycoprotein of the paramyxoviruses hendra virus (hev) and nipah virus (niv) have been discovered [corrected]. objective: to examine, compare and contrast the functional characteristics of hmabs with the potential for prophylaxis and treatment of diseases caused by sars cov, hev and niv. methods: ... | 2009 | 19216624 |
| case definitions for the 4 diseases requiring notification to who in all circumstances under the ihr (2005). | | 2009 | 19219965 |
| interaction between sars-cov helicase and a multifunctional cellular protein (ddx5) revealed by yeast and mammalian cell two-hybrid systems. | to reveal the putative cellular factors involved in sars coronavirus replication, the helicase (hel, nsp13) of sars coronavirus was used to screen the cdna library of rat pulmonary epithelial cells using the yeast two-hybrid system. positively interacting proteins were further tested using a mammalian cell hybrid system and co-immunoprecipitation in the human a549 cell line, which has been shown to support sars coronavirus replication. out of the seven positive clones observed by yeast two-hybri ... | 2009 | 19224332 |
| aryl diketoacids (adk) selectively inhibit duplex dna-unwinding activity of sars coronavirus ntpase/helicase. | as anti-hcv aryl diketoacids (adk) are good metal chelators, we anticipated that adks might serve as potential inhibitors of sars cov (scv) ntpase/helicase (hel) by mimicking the binding modes of the bismuth complexes which effectively competes for the zn(2+) ion binding sites in scv hel thereby disrupting and inhibiting both the ntpase and helicase activities. phosphate release assay and fret-based assay of the adk analogues showed that the adks selectively inhibit the duplex dna-unwinding acti ... | 2009 | 19233643 |
| differential characteristics of the early stage of lung inflammation induced by sars-cov nucleocapsid protein related to age in the mouse. | severe acute respiratory syndrome (sars) is an acute infectious disease of the respiratory system which has newly emerged. interestingly, it appears to be a disease that predominantly affects adults while the mortality in children is extremely low. however, the pathogenesis of sars in relation to different characteristics relevant to age remains unclear. | 2009 | 19234811 |
| [expression, purification and antibody preparation of recombinat sars-cov x5 protein]. | x5 protein is one of the putative unknown proteins of sars-cov. the recombinant protein has been successfully expressed in e. coli in the form of insoluble inclusion body. the inclusion body was dissolved in high concentration of urea. affinity chromatography was preformed to purify the denatured protein, and then the product was refolded in a series of gradient solutions of urea. the purified protein was obtained with the purity of > 95% and the yield of 93.3 mg x l(-1). polyclonal antibody of ... | 2008 | 19239038 |
| detection of novel sars-like and other coronaviruses in bats from kenya. | diverse coronaviruses have been identified in bats from several continents but not from africa. we identified group 1 and 2 coronaviruses in bats in kenya, including sars-related coronaviruses. the sequence diversity suggests that bats are well-established reservoirs for and likely sources of coronaviruses for many species, including humans. | 2009 | 19239771 |
| survival of surrogate coronaviruses in water. | the emergence of a previously unknown coronavirus infection, severe acute respiratory syndrome (sars), demonstrated that fecally contaminated liquid droplets are a potential vehicle for the spread of a respiratory virus to large numbers of people. to assess potential risks from this pathway, there is a need for surrogates for sars coronavirus to provide representative data on viral survival in contaminated water. this study evaluated survival of two surrogate coronaviruses, transmissible gastroe ... | 2009 | 19246070 |
| antibody fragment expression and purification. | interest in the potential of monoclonal antibodies (mabs) to serve as therapeutic agents has surged in the past decade with a major emphasis on human viral diseases. there has been much attention in this area directed towards the human immunodeficiency virus type-1 (hiv-1) and promising research developments have emerged on the inhibition of hiv-1 infection by mabs and the identification of several highly conserved neutralizing epitopes. more recently, potent fully-human neutralizing mabs have b ... | 2009 | 19252844 |
| thermostability of the n-terminal rna-binding domain of the sars-cov nucleocapsid protein: experiments and numerical simulations. | differential scanning calorimetry, circular dichroism spectroscopy, nuclear magnetic resonance spectroscopy, and numerical simulations were used to study the thermostability of the n-terminal rna-binding domain (rbd) of the sars-cov nucleocapsid protein. the transition temperature of the rbd in a mixing buffer, composed of glycine, sodium acetate, and sodium phosphate with 100 mm sodium chloride, at ph 6.8, determined by differential scanning calorimetry and circular dichroism, is 48.74 degrees ... | 2009 | 19254548 |
| mouse studies of sars coronavirus-specific immune responses to recombinant replication-defective adenovirus expressing sars coronavirus n protein. | 1. a recombinant adenovirus encoding sars coronavirus(sars-cov) nucleocapsid protein (rad-n) was constructed. 2. the ability of the rad-n to induce anti-sars-cov n antibody production and cellular immune responses was evaluated in an hla-a2.1/kb transgenic mouse model. | 2009 | 19258632 |
| roles of spike protein in the pathogenesis of sars coronavirus. | 1. infection with sars coronavirus (sars-cov) induces a cellular stress condition known as the unfolded protein response (upr). upr induction is mediated primarily by viral spike (s) protein. the modulation of upr by s protein involves activation of perk protein kinase. other branches of the upr pathways controlled by ire1 and atf6 proteins, respectively, are not involved. 2. the protease inhibitor ben-hcl effectively suppresses sars-cov infection by blocking virus entry. viral infectivity is as ... | 2009 | 19258633 |
| an oral mucosal dna vaccine for sars coronavirus infections. | 1. when different forms of sars coronavirus (sars-cov) spike protein-based vaccines for generation of a neutralising antibody response to sars-cov were injected into a mouse model, all the mice immunised with intramuscular tpa-optimised 800 dna vaccine boosted with intraperitoneal recombinant spike polypeptide generated by escherichia coli and intramuscular ctla4hinge sars800 dna vaccine boosted with intraperitoneal s-peptide had neutralising antibody titres of>1:1280.2. this observation may hav ... | 2009 | 19258634 |
| differential stepwise evolution of sars coronavirus functional proteins in different host species. | sars coronavirus (sars-cov) was identified as the etiological agent of sars, and extensive investigations indicated that it originated from an animal source (probably bats) and was recently introduced into the human population via wildlife animals from wet markets in southern china. previous studies revealed that the spike (s) protein of sars had experienced adaptive evolution, but whether other functional proteins of sars have undergone adaptive evolution is not known. | 2009 | 19261195 |
| comparison of vesicular stomatitis virus pseudotyped with the s proteins from a porcine and a human coronavirus. | the surface proteins s of severe acute respiratory syndrome coronavirus (sars-cov) and transmissible gastroenteritis virus (tgev) were compared for their ability to mediate infection of viral pseudotypes based on vesicular stomatitis virus (vsv). the cell tropism of the respective pseudotypes corresponded to the tropism of the viruses from which the s protein was derived. higher infectivity values were obtained with the sars-cov s protein than with the tgev s protein. differences were observed w ... | 2009 | 19264610 |
| suppression of host gene expression by nsp1 proteins of group 2 bat coronaviruses. | nsp1 protein of severe acute respiratory syndrome coronavirus (sars-cov), a group 2b cov, suppresses host gene expression by promoting host mrna degradation and translation inhibition. the present study analyzed the activities of nsp1 proteins from the group 2 bat cov strains rm1, 133, and hku9-1, belonging to groups 2b, 2c, and 2d, respectively. the host mrna degradation and translational suppression activities of nsp1 of sars-cov and rm1 nsp1 were similar and stronger than the activities of th ... | 2009 | 19264783 |
| drug targets in severe acute respiratory syndrome (sars) virus and other coronavirus infections. | coronaviruses are important human and animal pathogens of the order nidovirales. several new members were discovered following the emergence of sars-cov in human populations, including two human coronaviruses and several animal coronaviruses. they cause respiratory and gastrointestinal illnesses and have been found in the brains of patients with multiple sclerosis. the high mortality of sars, the identification of a natural reservoir, and the well-founded fear of provoking antibody-enhanced dise ... | 2009 | 19275708 |
| timely identification of optimal control strategies for emerging infectious diseases. | health authorities must rely on quarantine, isolation, and other non-pharmaceutical interventions to contain outbreaks of newly emerging human diseases. | 2009 | 19289133 |
| neutralizing antibody against severe acute respiratory syndrome (sars)-coronavirus spike is highly effective for the protection of mice in the murine sars model. | we evaluated the efficacy of three sars vaccine candidates in a murine sars model utilizing low-virulence pp and sars-cov coinfection. vaccinated mice were protected from severe respiratory disease in parallel with a low virus titer in the lungs and a high neutralizing antibody titer in the plasma. importantly, the administration of spike protein-specific neutralizing monoclonal antibody protected mice from the disease, indicating that the neutralization is sufficient for protection. moreover, a ... | 2009 | 19291090 |
| differential virological and immunological outcome of severe acute respiratory syndrome coronavirus infection in susceptible and resistant transgenic mice expressing human angiotensin-converting enzyme 2. | we previously reported that transgenic (tg) mice expressing human angiotensin-converting enzyme 2 (hace2), the receptor for severe acute respiratory syndrome coronavirus (sars-cov), were highly susceptible to sars-cov infection, which resulted in the development of disease of various severity and even death in some lineages. in this study, we further characterized and compared the pathogeneses of sars-cov infection in two of the most stable tg lineages, ac70 and ac22, representing those suscepti ... | 2009 | 19297479 |
| biochemical characterization of arterivirus nonstructural protein 11 reveals the nidovirus-wide conservation of a replicative endoribonuclease. | nidoviruses (arteriviruses, coronaviruses, and roniviruses) are a phylogenetically compact but diverse group of positive-strand rna viruses that includes important human and animal pathogens. nidovirus rna synthesis is mediated by a cytoplasmic membrane-associated replication/transcription complex that includes up to 16 viral nonstructural proteins (nsps), which carry common enzymatic activities, like the viral rna polymerase, but also unusual and poorly understood rna-processing functions. of t ... | 2009 | 19297500 |
| the 8ab protein of sars-cov is a luminal er membrane-associated protein and induces the activation of atf6. | the 8ab protein of sars-cov is a group-specific accessory protein, which is lost when the virus was transmitted from animals to humans due to a 29-nucleotide deletion in the orf8ab region. here we found that 8ab protein is associated with er membrane at luminal surface. 8ab protein was found to up-regulate the synthesis of endogenous er-resident chaperons involved in protein folding through the activation of the transcription factor atf6, while it showed no effect on the chop induction and xbp1 ... | 2009 | 19304306 |
| [prokaryotic expression of s2 extracellular domain of sars coronavirus spike protein and its fusion with hela cell membrane]. | to construct the expression plasmid of s2 extracellular domain (s2ed) of sars-coronavirus (sars- cov) spike protein (s protein) and enhanced green fluorescent protein (egfp) to obtain the fusion protein expressed in prokaryotic cells. | 2009 | 19304506 |
| c-terminal domain of sars-cov main protease can form a 3d domain-swapped dimer. | sars coronavirus main protease (m(pro)) plays an essential role in the extensive proteolytic processing of the viral polyproteins (pp1a and pp1ab), and it is an important target for anti-sars drug development. we have reported that both the m(pro) c-terminal domain alone (m(pro)-c) and the n-finger deletion mutant of m(pro) (m(pro)-delta7) exist as a stable dimer and a stable monomer (zhong et al., j virol 2008; 82:4227-4234). here, we report structures of both m(pro)-c monomer and dimer. the st ... | 2009 | 19319935 |
| activation of the sars coronavirus spike protein via sequential proteolytic cleavage at two distinct sites. | the coronavirus spike protein (s) plays a key role in the early steps of viral infection, with the s1 domain responsible for receptor binding and the s2 domain mediating membrane fusion. in some cases, the s protein is proteolytically cleaved at the s1-s2 boundary. in the case of the severe acute respiratory syndrome coronavirus (sars-cov), it has been shown that virus entry requires the endosomal protease cathepsin l; however, it was also found that infection of sars-cov could be strongly induc ... | 2009 | 19321428 |
| expression and membrane integration of sars-cov e protein and its interaction with m protein. | the severe acute respiratory syndrome (sars)-cov e gene fragment was cloned and expressed as a recombinant protein fused with a myc tag at the n-terminus in vitro and in vero e6 cells. similar to other n-glycosylated proteins, the glycosylation of sars-cov e protein occurred co-translationally in the presence of microsomes. the sars-cov e protein is predicted to be a double-spanning membrane protein lacking a conventional signal peptide. both of the transmembrane regions (a.a. 11-33 and 37-59) a ... | 2009 | 19322648 |
| structural insights into immune recognition of the severe acute respiratory syndrome coronavirus s protein receptor binding domain. | the spike (s) protein of the severe acute respiratory syndrome coronavirus (sars-cov) is responsible for host cell attachment and fusion of the viral and host cell membranes. within s the receptor binding domain (rbd) mediates the interaction with angiotensin-converting enzyme 2 (ace2), the sars-cov host cell receptor. both s and the rbd are highly immunogenic and both have been found to elicit neutralizing antibodies. reported here is the x-ray crystal structure of the rbd in complex with the f ... | 2009 | 19324051 |
| binding of sars coronavirus to its receptor damages islets and causes acute diabetes. | multiple organ damage in severe acute respiratory syndrome (sars) patients is common; however, the pathogenesis remains controversial. this study was to determine whether the damage was correlated with expression of the sars coronavirus receptor, angiotensin converting enzyme 2 (ace2), in different organs, especially in the endocrine tissues of the pancreas, and to elucidate the pathogenesis of glucose intolerance in sars patients. the effect of clinical variables on survival was estimated in 13 ... | 2010 | 19333547 |