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proteolytic activity and general characteristics of a marine bacterium, aeromonas proteolytica sp. n.merkel, joseph r. (fort johnson marine biological laboratory, college of charleston, charleston, s.c.), eugene d. traganza, barid b. mukherjee, travis b. griffin, and j. m. prescott. proteolytic activity and general characteristics of a marine bacterium, aeromonas proteolytica sp. n. j. bacteriol. 87:1227-1233. 1964.-a highly proteolytic bacterial species was isolated from the alimentary canal of the marine borer, limnoria. the morphological and biochemical characteristics of the organism indica ...19645874543
partial purification and some enzymatic properties of a proteinase from aeromonas proteolytica. 19695787115
[aeromonas hydrophila subsp. proteolytica (merkel et al. 1964) comb. nov]. 19695386248
regulation of proteolytic enzyme production by aeromonas proteolytica. i. extracellular endopeptidase. 19705415964
regulation of proteolytic enzyme production by aeromonas proteolytica. ii. extracellular aminopeptidase. 19705415965
some physical characteristics of a proteinase from aeromonas proteolytica. 19705442824
effect of poly i:c on parainfluenza 3 virus multiplication in wish cells. 19714396418
effects of antibiotics on the development and stability of mitochondrial enzymes in saccharomyces cerevisiae. 19724343088
virus infections of horses at newmarket, 1972 and 1973. 19744375339
chromocentres in polymorphs as interphase markers for chromosomes having increased constitutive heterochromatin. 19744140910
aeromonas proteolytica--a halophilic aeromonad?aeromonas proteolytica, a halophilic bacterium originally isolated from the intestine of a marine isopod, was compared with 93 strains of aeromonas hydrophila and representative strains of five similar genera in a computer and serological analysis. aeromonas proteolytica was peritrichate and non-reactive with six a. hydrohila antisera, and possessed a percentage guanine + cytosine ratio (49.5) outside the range for other aeromonads (55-62%). moreover, in the computer analysis it shared a lower l ...19751148936
[salmonellosis carrier state]. 19751214656
organ cultures of rat and hamster colon.human fibrin foam as a matrix for three-dimensional organ cultures was successfully employed for the cultivation of newborn rat and hamster colon tissue. colonic tissue was maintained under different oxygen atmospheres and with various synthetic media. explants of rat and hamster colon, maintained in leibovitz's l-15 medium buffered with hepes in the presence of 5% co2 and air, retained their normal columnar epithelial architecture for 24 days in culture. outgrowth of epithelial cells from colon ...1975165141
esterase activity of zinc neutral proteases.the hydrolysis of a series of depsipeptides demonstrates that the zinc neutral endopeptidases of bacteria are active esterases. esters such as bzgly-ophe-ala, bzgly-oleu-ala, and fa-gly-oleu-nh2 are hydrolyzed at rates three- to eightfold slower than are their exact peptide analogues, when hydrolyzed by thermolysin, bacillus subtilis neutral protease and the neutral protease from aeromonas proteolytica. ester hydrolysis by zinc neutral proteases follows the characteristic preference for hydropho ...19762276
superactivation of neutral proteases: acylation with n-hydroxysuccinimide esters.a series n-hydroxysuccinimide esters of acylamino acids previously shown to acylate and thereby increase the activity of thermolysin by several orders of magnitude (blumberg, s., and vallee, b. l. (1975), biochemistry 14, 2410) has been used to modify the related neutral proteases from bacillus subtilis, bacillus megaterium, and aeromonas proteolytica. each of these enzymes is activated to a level characteristic of the particular protein and the particular acyl group incorpporated when monitored ...1976823965
quantitative characteristics of the feyrter (apud) cells of the neonatal rabbit lung in normoxia and chronic hypoxia.our studies show that the apparent number of feyrter cells in the lung declines during the neonatal period in normoxic rabbits, and that in hypoxic animals a uniformly and significantly lower number of cells occurs as compared with the normoxic rabbits. there is some indication of degranulation of cells in the hypoxic groups. it is suggested that environmental and/or physiological factors associated with the start of extrauterine life, or lung development, may affect the apparent number and prob ...197722136
[immunological relations between mother and child during pregnancy (author's transl)].the relations between the maternal and foetal organisms during pregnancy are partly very complicated and remain inexplicable. blastocyst, trophoblast, embryo and foetus have membrane antigens at a very early stage, and the immunocompetence of the foetus develops already during the second third of the pregnancy period. many non-specific factors are involved in preventing the castingoff reaction of the foetal organism. the trophoblastic tissue is probably of central importance. the focal point of ...197898927
effects of morphine withdrawal on food competition hierarchies and fighting behavior in rats. 1979106426
interaction of amino acids with glycl-l-leucine hydrolysis and transport in monkey small intestine.1. there are two saturable transport processes in the monkey small intestine for glycyl-l-leucine, one with vmax. 1 mumol min-1 g-1 wet weight of tissue and an affinity constant (kt) of 5 mmol/l, and the other with vmax 3.9 mumol min-1 g-1 wet weight of tissue and kt 33 mmol/l. 2. glycyl-l-leucine uptake is inhibited by a wide variety of amino acids, although to a variable extent. the inhibition was shown to be competitive with leucine used as the representative amino acid. phenylalanine, methio ...1979117966
evidence for the subcellular localization and specificity of chlordane inhibition in the marine bacterium aeromonas proteolytica.sublethal levels (10 to 100 micrograms/ml) of the chlorinated insecticide chlordane (1,2,4,5,6,7,8,8-octachloro-3a,4,7,7a-tetrahydro-4,7-methanoindan) were introduced into the growth medium of the marine bacterium, aeromonas proteolytica. chlordane inhibited the synthesis of an extracellular endopeptidase by almost 40% but exhibited no such inhibition of the extracellular aminopeptidase also produced during the growth cycle. studied with 14c-labeled chlordane demonstrated that the insecticide wa ...1979156517
response to immunological or cholinergic stimulation of isolated mast cells from man, guinea pig and rat. 197991954
[cutaneous leishmaniasis. alcoy: an endemic zone]. 197992178
[effect of ergoline derivatives on platelet and blood vessels].the influence of dihydrogenated ergot alkaloids on vessels and blood platelets was studied in vitro and in vivo. at low concentrations they cause contraction of isolated vein strips via a stimulation of alpha-adrenoceptors, at higher concentrations they antagonize the action of noradrenaline on arteries and veins and inhibit the adrenaline-induced platelet aggregation in vitro. in volunteers, intravenous and subcutaneous administration of dihydroergotamine (dihytamin) caused a decrease in blood ...197994864
physiological responses of bacteria to cytochalasin a: effects on growth, transport, and enzyme induction.cytochalasin a at 5 to 25 microgram/ml (1.0 x 10(-5) to 5.2 x 10(-5) m) inhibited the growth of three gram-positive bacteria, arthrobacter sialophilus, staphyloccus aureus, and bacillus amyloliquifaciens, but had little or no effect on the growth of three gram-negative bacteria, excherichia coli, pseudomonas maltophilia, and aeromonas proteolytica. a. sialophilus and s. aureus recovered spontaneously from cytochalasin a-mediated growth inhibition after a considerable lag period, which was depend ...1979422516
managing metastatic breast cancer: a point of view. 1979218377
stereospecificity of amino acid hydroxamate inhibition of aminopeptidases.hydroxamates of amino acids and aliphatic acids are effective inhibitors of aeromonas proteolytica amino-peptidase (ec 3.4.11.10) and of both the cytosolic (ec 3.4.11.1) and microsomal (ec 3.4.11.2) aminopeptidases of swine kidney. cytosolic leucine aminopeptidase and the aeromonas enzyme were inhibited to a greater extent by d isomers than by the l enantiomorphs, manganese-activated kidney cytosolic leucine aminopeptidase being inhibited 10 times more effectively by d-leucine and d-valine hydro ...19836643439
immunodominant protein epitopes. i. induction of suppression to hen egg white lysozyme is obliterated by removal of the first three n-terminal amino acids.the lack of response to hen egg white lysozyme (hel) by c57bl (h-2b) mice has been demonstrated previously to be related to the induction of suppressor t (ts) cells which recognize the amino terminal region of hel. in this report, the nature of the protein determinant required for ts cell induction is more precisely detailed using des-1,2,3-hel (ap-hel) prepared with an aminopeptidase purified from aeromonas proteolytica . remarkably, the removal of just these three amino acids obliterates the a ...19846202528
conversion of estrogen receptor from a state with low affinity for estradiol into a state of higher affinity does not require 4s to 5s dimerization.this study was undertaken to establish whether the heat-promoted conversion of receptor-estradiol complex (re2) from a state with fast into a state with slow e2 dissociation requires 8s/4s to 5s transformation. the calf uterine estrogen receptor labeled with [3h]e2 at 0 c (state with low affinity for e2) was immobilized on hydroxylapatite (hap) in the absence (8s oligomer) or presence (4s monomer) of 0.4 m kcl and heated at 28 c in the presence of unlabeled diethylstilbestrol. under these condit ...19852981069
[role of virus-induced autoreactive t-lymphocytes, t-suppressors and the serum factor regulating their activity in the pathogenesis of experimental infection caused by the langat virus in mice].the precursors of autoreactive t-lymphocytes (partl) have been detected in the spleen of mice infected with langat virus. when introduced into syngeneic recipients, partl differentiate in their lymph nodes into autoreactive t-lymphocytes (artl) causing a fatal autoimmune disease in the syngeneic recipients in vivo and capable of destroying syngeneic cell cultures in vitro. in the thymus of mice infected with langat virus t-suppressors (ts) inhibiting the differentiation of partl into artl have b ...19852998127
effect of ultraviolet radiation on guinea pig epidermal langerhans cell cytomembrane: light and electron microscopic studies.the epidermal langerhans cells (elc) are thought to be the outermost station of the immune system. their location in the suprabasal layer of the epidermis makes them susceptible to a continuous exposure to ultraviolet irradiation (uvr) and if uvr alters the elc, this may in turn induce both local and systemic effects. in this paper we have shown that uva and uvb exposure produced dose-dependent changes of elc, characterized by disturbance of spatial distribution and aggregation of the atpase loc ...19853161009
induction of creatine kinase in immature rat uteri by zearalenone, an estrogenic mycotoxin.intraperitoneal administration of alpha-zearalenone (zen) to female immature rats induced synthesis of a uterine protein which has been identified as creatine kinase. the induced enzyme was purified to homogeneity by chromatography on deae sephacel and hydroxyapatite ultrogel. guinea pig antiserum against estrogen-induced uterine, rat uterine creatine kinase crossreacted with the zen-induced enzyme, indicating that zen exhibits an early estrogenic response in a manner analogous to natural estrog ...19853161514
enhanced replication of murine cytomegalovirus in murine leukemic lymphocytes.replication in vitro of murine cytomegalovirus was found to be enhanced in lymphoid cells from leukemic (akr/j) mice as compared with similar cells from nonleukemic animals. prolonged productive murine cytomegalovirus infection in lymphoid organs in vivo was demonstrable only in leukemic akr/j mice. latent nonproductive murine cytomegalovirus infection established in young nonleukemic akr/j mice was invariably reactivated and expressed in salivary glands and lymphoid organs after these animals b ...19862998605
issues relating to testing for hiv antibody (anti-hiv) 19863469567
[what is your diagnosis? what measures do you recommend?]. 19863775354
[dissecting microscopy observation of villous morphology in human normal duodenal epithelium]. 19863784079
lesions of the amygdala, but not of the cerebellum or red nucleus, block conditioned fear as measured with the potentiated startle paradigm.rats were given 10 light-shock pairings on 2 successive days. at 24-48 hr following training, groups of rats received bilateral transection of the cerebellar peduncles, bilateral lesions of the red nucleus (which receives most of the cerebellar efferents), or bilateral lesions of the central nucleus of the amygdala. control rats were sham operated. at 3-4 days after surgery, the rats were tested for potentiated startle (increased acoustic startle in the presence of the light previously paired wi ...19863954873
stimulation of bacterial dna synthesis by algal exudates in attached algal-bacterial consortia.algal-bacterial consortia attached to polystyrene surfaces were prepared in the laboratory by using the marine diatom amphora coffeaeformis and the marine bacterium vibrio proteolytica (the approved name of this bacterium is vibrio proteolyticus [w. e. c. moore, e. p. cato, and l. v. h. moore, int. j. syst. bacteriol. 35:382-407, 1985]). the organisms were attached to the surfaces at cell densities of approximately 5 x 10 cells cm (diatoms) and 5 x 10 cells cm (bacteria). the algal-bacterial con ...198616347218
a cytosolic protein contains a cryptic mitochondrial targeting signal.cytosolic dihydrofolate reductase from mouse contains a cryptic mitochondrial targeting sequence. if this sequence is attached to the amino terminus of 'passenger' proteins which by themselves cannot enter mitochondria, the resulting fusion proteins are transported into yeast mitochondria.19873543689
[bacteriologic status of the muscle tissue of broilers in the fattening phase and following meat removal]. 19873632600
the investigation and management of arthropod bite reactions acquired in the home. 19873119262
plant glycosides in a liposomal drug-delivery system.plant glycosides were incorporated into the liposomal surface to study their sugar-specific uptake by various tissues. two steroid glycosides, namely floribundasaponin d, with rhamnose as terminal sugar, and gracillin, with glucose and rhamnose as end sugars, were selected for the purpose. 125i-human igg encapsulated liposomes composed of egg lecithin (phosphatidylcholine), cholesterol, dicetyl phosphate (optional) and either floribundasaponin d or gracillin, when injected into the tail vein of ...19873426542
preparative electrochemical reduction of 2-amino-6-chloropurine and synthesis of 6-deoxyacyclovir, a fluorescent substrate of xanthine oxidase and a prodrug of acyclovir.d.c. polarography of 2-amino-6-chloropurine in aqueous medium over a broad ph range revealed two diffusion waves, the first of which corresponds to reduction of the c(6)-cl bond, leading to formation of 2-aminopurine in high yield. condensation of the sodium salt of 2-aminopurine with (2-acetoxyethoxy)methyl chloride led to the two isomeric 9- and 7-(2-hydroxyethoxymethyl)-2-aminopurines. the 9- isomer, 6-deoxyacyclovir, a prodrug of acyclovir previously synthesized by another route, was readily ...19872833054
two distinct mechanisms for regulation of gamma-glutamyl transpeptidase in cultured rat hepatocytes by glucocorticoid-like steroids.adult rat hepatocytes maintained in primary monolayer culture with defined medium were used to characterise two effects of glucocorticoid-like steroids in regulating gamma-glutamyltranspeptidase (ggt). low concentrations of glucocorticoids alone had little effect on ggt but synergistically enhanced induction of the enzyme by liver tumor-promoting xenobiotics such as 1,1,1-trichloro-2,2-bis-(4-chlorophenyl)-ethane (ddt) and hexachlorocyclohexane. the enhancing effect appears to be mediated by the ...19872885003
beta-adrenergic receptor agonist infusion increases plasma prostaglandin f levels in pregnant sheep.recent evidence of a stimulatory effect of beta-adrenergic receptor agonists on prostaglandin production by human fetal membranes in vitro prompted us to investigate whether prostaglandins are increased during beta-agonist infusion in vivo. on days 120 to 125 of gestation, blood samples were drawn from the aorta and vena cava of five crossbred ewes at 15-minute intervals for 1 hour before, during, and for 1 hour after a 3-hour infusion of isoproterenol (0.16 microgram/kg/min) or normal saline so ...19872890305
effects of neonatal 6-hydroxydopamine administration on different substance p-containing sensory neurones.rats were treated neonatally with 6-hydroxydopamine. this resulted in a greater than 93% depletion of noradrenaline content of all tissues examined in adult animals. there was a significant increase in the substance p content of semilunar ganglion, l5 dorsal root ganglion and of sciatic nerve. there was a corresponding increase in substance p synthesis as measured by axonal transport in the sciatic nerve. the noradrenaline content and ratio of noradrenaline: substance p in the iris of control an ...19872442004
evolution and development of the outer acrosomal membrane (oam) and evidence that acrosin-inhibitors are proteins of the oam.an antiserum to the purified porcine outer acrosomal membrane (oam) was raised in rabbits and the igg fraction isolated by ammonium sulphate precipitation and ion exchange chromatography. the antibodies reacted exclusively with the acrosomal cap of the sperm head as revealed by indirect immunofluorescence. in addition they cross-reacted not only with the acrosomal part of the spermatozoa of all mammalian species tested (bull, horse, rabbit, rat, mouse, hamster, mole, antelope, monkey, man) but a ...19872446533
late appearance of phospholipid platelet-activating factor and leukotriene b4 in human skin after repeated antigen challenge.inflammatory mediators were assessed in supernatants of chamber fluids from eight ragweed- or grass-sensitive subjects during antigen-induced cutaneous inflammatory responses. platelet activating factor (paf) accumulated at concentrations of 1 pm to 90 mumol/l in six of eight subjects beginning at 3 hours and continuing for 9 hours after antigen challenge. leukotriene b4 (ltb4) was detectable at cutaneous sites of antigen challenge in five of five subjects throughout the 9-hour period at levels ...19892538500
serological cross-reactivity among chlamydial strains in a family outbreak of psittacosis.three members of a family of nine persons contracted psittacosis with severe pneumonia, respiratory failure, delirium, hepatitis and renal involvement. a newly purchased cockatiel was probably the primary source of infection but person-to-person transmission is likely to have taken place between twin brothers who shared a bedroom, one of whom had no direct contact with birds. type-specific chlamydial serological tests identified the infecting agent as chlamydia psittaci. the highest titres in th ...19892674292
serological cross-reactivity among chlamydial strains in a family outbreak of psittacosis.three members of a family of nine persons contracted psittacosis with severe pneumonia, respiratory failure, delirium, hepatitis and renal involvement. a newly purchased cockatiel was probably the primary source of infection but person-to-person transmission is likely to have taken place between twin brothers who shared a bedroom, one of whom had no direct contact with birds. type-specific chlamydial serological tests identified the infecting agent as chlamydia psittaci. the highest titres in th ...19892674292
collagen accumulation by human mammary epithelial cells in primary culture. 19892720795
histamine concentration and ca2+ mobilization in arterial smooth muscle.smooth muscle contraction is dependent on ca2+ entry from the extracellular space or release from intracellular stores. the sensitivity of these ca2+ sources to agonist concentration was evaluated by measuring myoplasmic [ca2+] (as estimated by aequorin), myosin phosphorylation, and isometric stress in the swine carotid media. high histamine concentrations produced transient elevations in [ca2+] and phosphorylation with rapid generation of near maximal stress. lower histamine concentrations prod ...19892750885
differential adhesion of metastatic rat mammary carcinoma cells to organ-derived microvessel endothelial cells and subendothelial matrix.the in vitro adhesion rates of rat 13762nf mammary adenocarcinoma cell clones of different spontaneous metastatic potentials to cloned microvessel endothelial cell monolayers and their subendothelial extracellular matrix were investigated. in this system, high rates of adhesion of the cloned tumor cell lines to syngeneic target (lung) organ-derived subendothelial matrix correlated with spontaneous metastatic potential, whereas adhesion to the lung microvessel endothelial cell apical surfaces occ ...19892776962
immunofluorescence demonstration of digoxin in human myocardial tissue and isolated rat cardiac myocytes with monoclonal antibodies.the aim of the present investigation was to identify the morphological correlates of digoxin binding sites in human heart muscle tissue and isolated viable rat heart myocytes. cardiac glycoside linked to myocardial cells was demonstrated by monoclonal digoxin specific antibody and by fitc-conjugated anti-mouse immunoglobin serum. this versatile immunofluorescence method can be used in diagnostic and experimental studies of cardiac glycoside binding.19902227731
the unique stability of vibrio proteolyticus neutral protease under alkaline conditions affords a selective step for purification and use in amino acid-coupling reactions.a procedure is described for the purification of a neutral protease from fermentation broths of vibrio proteolyticus. the key feature of the purification scheme is the selective, irreversible inactivation of a contaminating exoenzyme, aminopeptidase, by alkali treatment, rather than removal of this enzyme by conventional chromatographic methods. fermentation broths or concentrates were brought to ph 11.5 to 11.7 by na2co3-naoh addition and incubated at 25 degrees c until aminopeptidase activity ...19902403247
tenascin expression in the human endometrium and in endometrial adenocarcinomas.to investigate the involvement of tenascin, an extracellular matrix glycoprotein, in epithelial growth and malignancy, its specific distribution pattern in the human uterus was examined immunohistochemically. during the proliferative phase of the menstrual cycle, this antigen was found as a sharp band around the endometrial glands. the immunoreactivity persisted until the early postovulatory phase of the menstrual cycle, but was not detectable in the glandular or stromal compartment during this ...19901694256
structure of the rabbit phospholamban gene, cloning of the human cdna, and assignment of the gene to human chromosome 6.we have isolated and characterized genomic dna clones encoding rabbit phospholamban. only a single gene for phospholamban was detected in the rabbit genome. the phospholamban gene of 13.2 kilobases contains only one 10.5-kilobase intron, which separates exonic sequences located in the 5'-untranslated region. two potential transcription initiation sites were mapped to 335 and 185 nucleotides upstream from the translation initiation site in the mrna or 239 and 89 nucleotides upstream from the exon ...19911828805
complexity and sequence identification of 24 rat v beta genes.twenty-four tcr v beta genes were cloned by anchored pcr from the lewis rat strain and identified by nucleotide and amino acid sequence comparisons to known mouse v beta genes. rat v beta genes exist in 17 single-member and 3 multimember subfamilies and exhibit 86 to 94 and 72 to 92% nucleotide and amino acid sequence similarities, respectively, to their mouse counterparts. a single rat gene, designated v beta 20, having no previously known mouse counterpart was identified; a closely related gen ...19911828824
genes coding for integration host factor are conserved in gram-negative bacteria.a genetic system for the selection of clones coding for integration host factor and hu homologs is described. we demonstrate that the hima and hip genes of serratia marcescens and aeromonas proteolytica can substitute for the escherichia coli genes in a variety of biological assays. we find that the sequence and genetic organization of the hima and hip genes of s. marcescens are highly conserved.19911917861
measurement of sensitivity to adriamycin in hybrid spheroids.using the newly developed hybrid spheroid system, we have been able to measure sensitivities to adriamycin in two sublines of lovo cells (a human colon adenocarcinoma line). hybrid spheroids are composed of coagglomerated lovo cells, and nonproliferating hela "feeder" cells, mimicking a population of minitumors. spheroids of uniform size, but containing different numbers of lovo cells, were incubated with various concentrations of adriamycin and after washing, plated for colony formation. binomi ...19911933483
characterization of a pollen-specific cdna from sunflower encoding a zinc finger protein.we have isolated, via differential hybridization screening of a floral cdna library from sunflower, a cdna clone that hybridizes to a 1100 nucleotide-long mrna found exclusively in mature pollen grains. the cdna encodes a 219 amino acid-long polypeptide containing two potential zinc fingers alternating with two basic domains. a similar organization is found in the erythroid-specific transcription factor eryf1 from chicken and its murine homolog gf-1. the c-terminus of the protein contains a sixf ...19921302629
towards a phylogeny of the genus vibrio based on 16s rrna sequences.the inter- and intrageneric relationships of the genus vibrio were investigated by performing a comparative analysis of the 16s rrnas of 10 species, including four pathogenic representatives. the results of immunological and 5s rrna studies were confirmed in that the genus is a neighboring taxon of the family enterobacteriaceae. with regard to the intrageneric structure, vibrio alginolyticus, vibrio campbellii, vibrio natriegens, vibrio harveyi, vibrio proteolyticus, vibrio parahaemolyticus, and ...19921371064
equine herpesvirus 1 glycoprotein d: mapping of the transcript and a neutralization epitope.studies with molecular and immunological techniques identified and mapped the transcript encoding glycoprotein d (gd) of equine herpesvirus 1 kya, as well as two continuous gd antigenic determinants. three mrna species of 5.5, 3.8, and 1.7 kb overlap the gd open reading frame and are transcribed from the dna strand encoding gd. northern (rna) blot hybridization with both dna clones and riboprobes, as well as s1 nuclease analyses, showed the 3.8-kb mrna to encode gd and to be synthesized as a lat ...19921383565
[effect of peptides of the tracheal mucosa and vascular wall in acute hyperoxic lung injury].it was demonstrated that peptides of the tracheal mucosa (ptm) and peptides of the vascular wall (pvw) produce a certain protective effect in hyperoxic lung injury. the peptides, (ptm to a greater measure) prolonged the survival of mice and inhibited the dynamics of the growth of the lung coefficient in rats. ptm reduced the formation of malonic dialdehyde and increased the activity of glucose-6-phosphate dehydrogenase in the lungs. morphometric studies showed that ptm produce a more marked inhi ...19921480431
isolation and characterization of the hupa gene coding for hu of aeromonas proteolytica. 19921508695
rapid purification of the aeromonas proteolytica aminopeptidase: crystallization and preliminary x-ray data.the heat-stable aminopeptidase from aeromonas proteolytica has been purified using two new procedures, with the aim of preparing large single crystals for x-ray analysis. in a first procedure, we tried to avoid any drastic conditions capable of inducing microheterogeneities in the protein sample. the enzyme was purified through two chromatographic steps based on hydrophobic interactions and ion exchange. in a second procedure a heat treatment of the protein to a temperature of 70 degrees c over ...19921550363
cloning, sequencing and expression of the gene encoding the extracellular neutral protease, vibriolysin, of vibrio proteolyticus.the gene (nprv), encoding the extracellular neutral protease, vibriolysin (nprv), of the gram- marine microorganism, vibrio proteolyticus, was isolated from a v. proteolyticus dna library constructed in escherichia coli. the recombinant e. coli produced a protease that co-migrated with purified neutral protease from v. proteolyticus on non-denaturing polyacrylamide gels, and that demonstrated enzymatic specificity towards the neutral protease substrate n-[3-(2-furyl)acryloyl]-l-alanylphenylalani ...19921551587
isolation of the leucine aminopeptidase gene from aeromonas proteolytica. evidence for an enzyme precursor.the leucine aminopeptidase of aeromonas proteolytica (ec 3.4.11.10) is a monomeric metalloenzyme having the capacity to bind two zn2+ atoms in the active site. structural information of this relatively small aminopeptidase that could illuminate the catalytic mechanism of the metal ions is lacking; hence, we have obtained sequences from the purified enzyme, cloned the corresponding gene, and expressed the recombinant protein in escherichia coli. the deduced primary amino acid sequence of this sec ...19921569090
cloning and nucleotide sequence of the vibrio proteolyticus aminopeptidase gene.the gene encoding the vibrio proteolyticus aminopeptidase was cloned and sequenced and its amino acid sequence was deduced. the gene encodes a 54 kda protein, larger than the previously reported size of 30 kda for the purified aminopeptidase. sequence alignments revealed a 43-45% homology with two other vibrio sp. extracellular proteinases.19921627651
preliminary evaluation of vibriolysin, a novel proteolytic enzyme composition suitable for the debridement of burn wound eschar.vibriolysin, a proteolytic enzyme secreted by the marine microorganism vibrio proteolyticus, was evaluated for its efficacy as an enzymatic debridement agent. initial in vitro experiments revealed that the protease readily hydrolyzed proteinaceous components of eschar (e.g., fibrin, elastin, and in particular, collagen). enzyme selectivity towards the digestion of denatured proteins but not viable elements was demonstrated in vitro and in vivo by treatment of full-thickness burn wounds with a po ...19938245109
production of authentic human proapolipoprotein a-i in escherichia coli: strategies for the removal of the amino-terminal methionine.several methods were compared with respect to the production of authentic, n-terminal methionine-free proapolipoprotein a-i in engineered escherichia coli bacteria. a first approach consisted of treating the purified methionylated recombinant protein with an amino-peptidase, purified from aeromonas proteolytica. a second series of strategies was based on the construction of proapo a-i encoding cassettes carrying built-in recognition sites suitable for specific in vitro cleavage of the products w ...19937763464
a prototype stable rna identification cassette for monitoring plasmids of genetically engineered microorganisms.a prototype stable rna identification cassette for monitoring genetically engineered plasmids carried by strains of escherichia coli has been developed. the cassette consists of a vibrio proteolyticus 5s ribosomal rna (rrna) gene surrounded by promoters and terminators from the rrnb operon of escherischia coli. the identifier rna is expressed and successfully processed so that approximately 30% of the 5s rrna isolated from either whole cells or 70s ribosomes is of the v. proteolyticus type. cell ...199311539559
crystal structure of aeromonas proteolytica aminopeptidase: a prototypical member of the co-catalytic zinc enzyme family.aminopeptidases specifically cleave the amino-terminal residue from polypeptide chains and are involved in the metabolism of biologically active peptides. the family includes zinc-dependent enzymes possessing either one or two zinc ions per active site. structural studies providing a detailed view of the metal environment may reveal whether the one-zinc and two-zinc enzymes constitute structurally and mechanistically distinct subclasses, and what role the metal ions play in the catalytic process ...19948087555
a novel approach for monitoring genetically engineered microorganisms by using artificial, stable rnas.further improvements in technology for efficient monitoring of genetically engineered microorganisms (gems) in the environment are needed. technology for monitoring rrna is well established but has not generally been applicable to gems because of the lack of unique rrna target sequences. in the work described herein, it is demonstrated that a deletion mutant of a plasmid-borne vibrio proteolyticus 5s rrna gene continues to accumulate to high levels in escherichia coli although it is no longer in ...19957487004
metal content and conformation of the metalloprotease from the marine sponge spheciospongia vesparia.we have recently purified a protease from the marine sponge spheciospongia vesparia. it consists of a single nonglycosylated polypeptide chain with a molecular weight of 29 600 and has one free thiol group. metal analysis revealed the presence of zinc at 2.02 +/- 0.05 g-atoms per mole of protein, as measured by atomic absorption spectroscopy. the circular dichroism spectrum in the far uv region (183-259 nm) indicates that the sponge protease contains appreciable amounts of beta sheet. this enzym ...19958528145
the structure of the aeromonas proteolytica aminopeptidase complexed with a hydroxamate inhibitor. involvement in catalysis of glu151 and two zinc ions of the co-catalytic unit.the structure of the complex of aeromonas proteolytica aminopeptidase, a two-zinc exopeptidase, with the inhibitor p-iodo-d-phenylalanine hydroxamate has been determined by x-ray crystallography. refinement of the structure, which includes 220 water molecules, using data at 0.80-0.23-nm resolution resulted in a crystallographic residual r value of 16%. the hydroxamate group adopts a planar conformation whereby the two oxygen atoms interact with the zinc ions. the n-hydroxyl group of the inhibito ...19968647077
aminopeptidase from streptomyces griseus: primary structure and comparison with other zinc-containing aminopeptidases.the aminopeptidase from streptomyces griseus is a calcium-activated metalloenzyme, which contains 2 mol tightly bound zinc/mol protein. this aminopeptidase rapidly hydrolyzes peptide bonds formed by n-terminal hydrophobic amino acids, such as leucine, methionine and phenylalanine. we have determined the complete primary structure of the protein, which contains 284 amino acid residues, yielding a molecular mass of 29723 da. a search in the swiss-prot database for sequence similarities revealed a ...19968665903
bacterial aminopeptidases: properties and functions.aminopeptidases are exopeptidases that selectively release n-terminal amino acid residues from polypeptides and proteins. bacteria display several aminopeptidasic activities which may be localised in the cytoplasm, on membranes, associated with the cell envelope or secreted into the extracellular media. studies on the bacterial aminopeptide system have been carried out over the past three decades and are significant in fundamental and biotechnological domains. at present, about one hundred bacte ...19968703509
identification of vibrio proteolyticus with a differential medium and a specific probe.a differential medium (vp8) and a specific probe, based on the variable region v3 of the 16s rrna gene, for the detection of vibrio proteolyticus are defined. the medium contains 8% nacl, which allows selective growth of moderately halophilic vibrio strains. d-sorbitol, as the main carbon source, differentiates the species that can ferment it by the ph indicators cresol red and bromothymol blue. v. proteolyticus and 8 of 418 strains studied grew on the medium and used the d-sorbitol, forming bri ...19968779607
cloning and genetic analysis of the vibrio cholerae aminopeptidase gene.the structural gene for the vibrio cholerae leucine aminopeptidase (lap) was cloned and sequenced. the cloned dna fragment contained a 1,503-bp open reading frame potentially encoding a 501-amino-acid polypeptide with a calculated molecular mass of 54,442 da. the deduced amino acid sequence of the entire protein showed high homology with the sequence of vibrio proteolyticus leucine aminopeptidase. the residues potentially involved in binding the zinc ions were completely conserved in the v. chol ...19968890197
sequence, overproduction and purification of vibrio proteolyticus ribosomal protein l18 for in vitro and in vivo studies.a strategy suggested by comparative genomic studies was used to amplify the entire vibrio proteolyticus (vp) gene for ribosomal protein l18. vp l18 and its flanking regions were sequenced and compared with the deduced amino acid (aa) sequences of other known l18 proteins. a 26-aa residue segment at the carboxy terminus contains many strongly conserved residues and may be critical for the l18 interaction with 5s rrna. this approach should allow rapid characterization of l18 from large numbers of ...19968996113
[increased camp concentration in parotid secretion in arterial hypertension: relationship to plasma-renin activity (author's transl)].hypertensives with normal and increased plasma-renin activity have an increased salivary excretion of cyclic adenosine monophosphate (camp) such as can be obtained in normal persons by stimulation of beta-adrenergic fibres. propanolol, as a beta-receptor blocker, reduces such increased camp levels in hypertensives to minimal levels. the results suggest that beta-receptors are stimulated in hypertensives with normal or increased renin activity.1996168050
streptomyces griseus aminopeptidase: x-ray crystallographic structure at 1.75 a resolution.the x-ray crystal structure of the enzyme streptomyces griseus aminopeptidase (sgap) has been determined in its double zinc form to 1.75 a resolution, in its apo-enzyme from (zinc removed) to 2.1 a resolution, and as a mercury replaced derivative to 2.1 a resolution. the structure solution was achieved by single isomorphous replacement with phasing from anomalous scattering (siras), followed by density modification with histogram matching. the protein consists of a central beta-sheet made up of ...19979048953
crystal structure of carboxypeptidase g2, a bacterial enzyme with applications in cancer therapy.carboxypeptidase g enzymes hydrolyze the c-terminal glutamate moiety from folic acid and its analogues, such as methotrexate. the enzyme studied here, carboxypeptidase g2 (cpg2), is a dimeric zinc-dependent exopeptidase produced by pseudomonas sp. strain rs-16. cpg2 has applications in cancer therapy: following its administration as an immunoconjugate, in which cpg2 is linked to an antibody to a tumour-specific antigen, it can enzymatically convert subsequently administered inactive prodrugs to ...19979083113
mechanistic studies on the aminopeptidase from aeromonas proteolytica: a two-metal ion mechanism for peptide hydrolysis.the aminopeptidase from aeromonas proteolytica (aap) is uncompetitively inhibited by fluoride ion at ph 8.0 with an inhibition constant (ki) of 30 mm. thus, fluoride inactivates aap only after substrate binding, and only a single fluoride ion binds to aap. on the other hand, chloride ion does not inhibit aap up to concentrations of 2 m at ph 8.0. the ph dependence of fluoride inhibition of aap was measured over the ph range 6.0-9.5. between ph values of 6.0 and 9.0, fluoride ion acts as a pure u ...19979100023
continuous spectrophotometric assay of peptide deformylase.a continuous assay for peptide deformylase has been developed using a formylated dipeptide, formyl-met-leu-p-nitroanilide, as substrate. removal of the formyl group by a peptide deformylase renders the dipeptide product, which contains a free nh2 terminus, a substrate for an aminopeptidase from aeromonas proteolytica. sequential hydrolysis of the dipeptide by the aminopeptidase releases a p-nitroaniline, which is monitored spectrophotometrically at 405 nm. this assay was applied to determine the ...19979234895
spectroscopically distinct cobalt(ii) sites in heterodimetallic forms of the aminopeptidase from aeromonas proteolytica: characterization of substrate binding.the co(ii)zn(ii)- and zn(ii)co(ii)-substituted derivatives of the aminopeptidase from aeromonas proteolytica (aap) were probed by epr spectroscopy. epr spectra of the high-spin s = 3/2 co(ii) ions in [cozn(aap)] and [znco(aap)] indicated that each metal binding site provides a spectroscopically distinct signature. for [cozn(aap)], subtraction of epr spectra recorded at ph 7.5 and 10 revealed that two species were present and that the relative contributions to each of the experimental spectra wer ...19979245416
crystal structure at 1.2 a resolution and active site mapping of escherichia coli peptidyl-trna hydrolase.peptidyl-trna hydrolase activity from escherichia coli ensures the recycling of peptidyl-trnas produced through abortion of translation. this activity, which is essential for cell viability, is carried out by a monomeric protein of 193 residues. the structure of crystalline peptidyl-trna hydrolase could be solved at 1.2 a resolution. it indicates a single alpha/beta globular domain built around a twisted mixed beta-sheet, similar to the central core of an aminopeptidase from aeromonas proteolyti ...19979303320
a low molecular weight artificial rna of unique size with multiple probe target regions.artificial rnas (arnas) containing novel sequence segments embedded in a deletion mutant of vibrio proteolyticus 5s rrna have previously been shown to be expressed from a plasmid borne growth rate regulated promoter in e. coli. these arnas accumulate to high levels and their detection is a promising tool for studies in molecular microbial ecology and in environmental monitoring. herein a new construct is described which illustrates the versatility of detection that is possible with arnas. this 3 ...199711540055
fetal heart rate during a maternal grand mal epileptic seizure.although maternal ingestion of antiepileptic drugs is strongly suspected of causing congenital defects, particularly oral clefts, the effect of epilepsy itself or a combined effect of drug intake and epilepsy have not been excluded as etiological factors. very little is known about fetal oxygenation during a maternal grand mal epileptic seizure. we describe two cases in which fetal heart rate was recorded during a maternal epileptic seizure during labor. the first fetus became clearly asphyctic ...1998106102
note: molecular cloning of chitinase genes from vibrio anguillarum and v. parahaemolyticus.chitinase genes from vibrio anguillarum kv9001 and v. parahaemolyticus atcc17802 were cloned into escherichia coli. open reading frames of chitinase genes from v. anguillarum (vac) and v. parahaemolyticus (vpc) are 1755 bp and 1890 bp, respectively. the deduced amino acid sequences of these genes have 71.6% identity. there are two consensus sequence regions in the vac and vpc proteins. the vac gene was highly prevalent in v. anguillarum, and the dna probe of the vac gene hybridized to v. alginol ...19989717305
inhibition of streptomyces griseus aminopeptidase and effects of calcium ions on catalysis and binding--comparisons with the homologous enzyme aeromonas proteolytica aminopeptidase.streptomyces griseus aminopeptidase is a zinc metalloenzyme containing 2 mol zinc/mol protein, similar to the homologous enzyme aeromonas proteolytica aminopeptidase. in addition, a unique ca2+-binding site has been identified in the streptomyces enzyme, which is absent in the aeromonas enzyme. binding of ca2+ enhances stability of the streptomyces enzyme and modulates its activity and affinity towards substrates and inhibitors in a structure-dependent manner. among the three hydrophobic 4-nitro ...19989874195
site-directed mutagenesis of predicted active site residues in glutamate carboxypeptidase ii.glutamate carboxypeptidase ii (gcp ii) catalyzes the extracellular hydrolysis of the neuromodulator n-acetyl-aspartylglutamate to n-acetyl-aspartate and glutamate. gcp ii also hydrolyzes gamma-glutamyl bonds in folylpolyglutamate. the predicted amino acid sequence of gcp ii displays similarities to aminopeptidases from streptomyces griseus and vibrio proteolyticus, whose crystal structures have been determined. these aminopeptidases are cocatalytic zinc metallopeptidases belonging to the peptida ...19999882712
thermostable aminopeptidase from pyrococcus horikoshii.from the genome sequence data of the thermophilic archaeon pyrococcus horikoshii, an open reading frame was found which encodes a protein (332 amino acids) homologous with an endoglucanase from clostridium thermocellum (42% identity), deblocking aminopeptidase from pyrococcus furiosus (42% identity) and an aminopeptidase from aeromonas proteolytica (18% identity). this gene was cloned and expressed in escherichia coli, and the characteristics of the expressed protein were examined. although endo ...199910218575
lapstatin, a new aminopeptidase inhibitor produced by streptomyces rimosus, inhibits autogenous aminopeptidases.lapstatin, a low-molecular-weight aminopeptidase inhibitor, was purified to homogeneity from streptomyces rimosus culture filtrates. the purification procedure included extraction with methanol, followed by chromatography on dowex 50wx4, ag50wx4, and hplc rp c18 columns. by amino acid analysis, mass spectrometry, and nmr spectroscopy, the structure of lapstatin was shown to be 3-amino-2-hydroxy-4-methylpentanoylvaline. lapstatin inhibited the extracellular leucine aminopeptidases from streptomyc ...199910369895
recombinant light chain of human monoclonal antibody hb4c5 as a potentially useful lung cancer-targeting vehicle.recombinant lambda light chain of lung cancer-reacting human monoclonal antibody hb4c5 was expressed in escherichia coli. expression in bacteria ensured the generation of homogeneous light chain species devoid of activity-hampering n-linked glycosylation usually found in the light chain cdr-1 of hb4c5. molecular engineering was also employed to eliminate the c-terminal two amino acid residues, i.e., cys and ser, to prevent the formation of lambda light chain dimers which are less reactive than t ...199910405832
1-butaneboronic acid binding to aeromonas proteolytica aminopeptidase: a case of arrested development.hydrolases containing two metal ions connected by a bridging ligand catalyze reactions important in carcinogensis, tissue repair, post-translational modification, control and regulation of biochemical pathways, and protein degradation. the aminopeptidase from aeromonas proteolytica serves as a paradigm for the study of such bridged bimetallic proteases since its three-dimensional structure is known to very high resolution and its catalytic reaction is amenable to spectroscopic examination. herei ...199910413478
inhibition of the aminopeptidase from aeromonas proteolytica by aliphatic alcohols. characterization of the hydrophobic substrate recognition site.seven aliphatic and two aromatic alcohols were tested as reporters of the substrate selectivity of the aminopeptidase from aeromonas proteolytica (aap). this series of alcohols was chosen to systematically probe the effect of carbon chain length, steric bulk, and inhibitor shape on the inhibition of aap. initially, however, the question of whether aap is denatured in the presence of aliphatic alcohols was addressed. on the basis of circular dichroism (cd), electronic absorption, and fluorescence ...199910471294
cold-adapted alanine dehydrogenases from two antarctic bacterial strains: gene cloning, protein characterization, and comparison with mesophilic and thermophilic counterparts.the genes encoding nad(+)-dependent alanine dehydrogenases (aladhs) (ec 1.4.1.1) from the antarctic bacterial organisms shewanella sp. strain ac10 (shealadh) and carnobacterium sp. strain st2 (caraladh) were cloned and expressed in escherichia coli. of all of the aladhs that have been sequenced, shealadh exhibited the highest level of sequence similarity to the aladh from the gram-negative bacterium vibrio proteolyticus (vpraladh). caraladh was most similar to aladhs from mesophilic and thermoph ...199910473410
slow-binding inhibition of the aminopeptidase from aeromonas proteolytica by peptide thiols: synthesis and spectroscopic characterization.peptide-derived thiols of the general structure n-mercaptoacyl-leucyl-p-nitroanilide (1a-c) were synthesized and found to be potent, slow-binding inhibitors of the aminopeptidase from aeromonas proteolytica (aap). the overall potencies (k(i)) of these inhibitors against aap range from 2.5 to 57 nm exceeding that of the natural product bestatin and approaching that of amastatin. the corresponding alcohols (2a-b) are simple competitive inhibitors of much lower potencies (k(i) = 23 and 360 microm). ...199910569943
the extracellular regions of psma and the transferrin receptor contain an aminopeptidase domain: implications for drug design.the aeromonas proteolytica aminopeptidase (amp), pseudomonas sp. (rs-16) carboxypeptidase g2 (cpg2), and streptomyces griseus aminopeptidase (sgap) are zinc dependent proteolytic enzymes with cocatalytic zinc ion centers and a conserved aminopeptidase fold. a blast search with the sequence of the solved amp structure indicated that a similar domain could be found in prostate-specific membrane antigen (psma) and the transferrin receptor (tfr). when the psma or tfr sequence was input into the thre ...199910595564
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