Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0272170 (SDS)
 50,377 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Matrix metalloproteinases are thought to play major roles in a wide array of normal and pathological processes. These proteinases are involved in the degradation of the extracellular matrix and are believed to facilitate the movement of cells from one site to another. In the current study, we examined the expression of the 92 kDa gelatinase activity (MMP-9) by the human T-lymphoma cell line, HSB. Proteinase activity was greatly elevated when cells were treated with TPA. This induction was initially observed at 6 h post-TPA treatment and continued to increase up to 48 h. Proteinase induction was inhibited by actinomycin D and cycloheximide, indicating that nascent RNA and protein synthesis were required. Staurosporine, an inhibitor of protein kinase C activity, suppressed the TPA-induction of gelatinase activity. Our results suggest that TPA induces the 92 kDa gelatinase activity by activating protein kinase C. TGF-beta also induced proteinase activity, although to a lesser extent than TPA. Several criteria indicate that this enzyme is a member of the family of matrix metalloproteinases: (1) this activity was inhibited by EDTA, 1,10-phenanthroline and TIMP; (2) this activity bound to a gelatin-agarose affinity resin; (3) it has a mass of approx. 92 kDa on SDS-polyacrylamide gels; (4) it cleaves gelatin and (5) the inducible proteinase cross reacts with antiserum to MMP-9.
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PMID:Induction of metalloproteinase activity in human T-lymphocytes. 849 86

An extracellular proteinase produced by the filamentous fungus Scedosporium apiospermum has been purified and characterized. Initially, in vitro conditions for enzyme synthesis were investigated. The highest yield of enzyme production was obtained when the fungus was cultivated in modified Czapek-Dox liquid medium supplemented with 0.1% bacteriological peptone and 1% (w/v) glucose as the nitrogen and carbon sources respectively. Purification to homogeneity of the proteinase was accomplished by (NH4)2SO4 precipitation, followed by gel filtration through Sephadex G-75 and finally affinity chromatography through immobilized phenylalanine. Analysis of the purified enzyme by SDS/PAGE revealed a single polypeptide chain with an apparent molecular mass of 33 kDa. Further investigation of its physical and biochemical properties disclosed numerous similarities with those of the previously described serine proteinase of Aspergillus fumigatus. The enzyme was not glycosylated and its pI was 9.3. Proteinase activity was optimum between 37 and 50 degrees C and at pH 9.0, but remained high within a large range of pH values between 7 and 11. The inhibition profile and N-terminal amino acid sequencing confirmed that this enzyme belongs to the subtilisin family of serine proteinases. In agreement with this, the specific synthetic substrate N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide proved to be an excellent substrate for the proteinase with an estimated Km of 0.35 mM. Like the alkaline proteinase of A. fumigatus, this enzyme was able to degrade human fibrinogen, and thus may act as a mediator of the severe chronic bronchopulmonary inflammation from which cystic fibrosis patients suffer.
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PMID:A 33 kDa serine proteinase from Scedosporium apiospermum. 867 95

We have isolated the cDNA and the genomic clones encoding a novel serine proteinase, named proteinase T, from the fungus Tritirachium album Limber. The coding region of the gene is interrupted by two introns. The amino acid sequence of proteinase T as deduced from the nucleotide sequence is about 56% identical to that of proteinase K. Four cysteines are present in the mature proteinase, probably in the form of disulfide bonds. We have also purified the native proteinase from Tritirachium album Limber grown in the presence of 2% skim milk. Proteinase T is extremely stable at 50 degrees C. The thermal stability is not affected in the presence of 1% SDS either at pH 8.0 or 10.0. We have expressed the cDNA of proteinase T in Escherichia coli. The authenticity of the proteinase has been characterized by Western blotting and amino terminal analysis of the recombinant product. High level expression of proteinase T in E. coli as well as the refolding process to generate active proteinase will be discussed in detail.
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PMID:Cloning and expression of the gene encoding a novel proteinase from Tritirachium album limber. 879 13

Characterization of a purified proteinase from Trichomonas vaginalis was carried out using bacitracin-sepharose affinity chromatography. Trichomonas vaginalis KT-9 isolate was used as a source of enzyme study. Proteinase activity was determined using Bz-Pro-Phe-Arg-Nan as the substrate. Optimum pH for the purified proteinase activity was 7.0 and 6.0, 9.0 with DTT. Optimum temperature was 37 degrees C and isoelectric point was 7.2. Activity of this proteinase was inhibited by E-64, antipain, leupeptin, Hg2+ and Zn2+ and activated by DTT and cysteine. Activity of the purified proteinase was visualized by gelatin SDS-PAGE. The gelatinolytic activity of the purified proteinase was inhibited by E-64, antipain, leupeptin, and IAA, but not by PMSF and EDTA. On SDS-PAGE, the molecular weight of the purified proteinase was 60,000 daltons. Sera of rabbits infected with T. vaginalis reacted specifically in immunoblots with this proteinase. These results indicate that 60 kDa of purified proteinase was cysteine proteinase with antigenicity.
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PMID:[Characterization of the partially purified proteinase from Trichomonasvaginalis]. 882 Jul 41

Proteinase K-digested cell lysates from 25 Campylobacter fetus subspecies fetus and C. fetus subsp. venerealis strains were examined by SDS-PAGE and immunoblotting. Three SDS-PAGE lipopolysaccharide (LPS) profiles were observed. Two profiles were consistent with those previously reported for serogroup A and serogroup B and AB isolates and were distinguished by the relative mobility of bands in the O-chain region and by a strong reaction on immunoblots with homologous antisera. The third profile was similar but had faster migrating O-chain bands. Immunoblot reactions using homologous and heterologous adsorbed antisera showed that the O-antigen of the C. fetus subsp. fetus reference strain and other profile 2-type LPS strains was distinct from the O-antigens of strains with profile 1- or profile 3-type LPS. O-antigens of strains with profile 1- and profile 3-type LPS had shared epitopes. One strain had core components but no detectable O-antigens. Common core LPS antigens appear to be present in all strains and antibodies to common core LPS epitopes may be useful reagents for rapid detection of C. fetus.
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PMID:Electrophoretic and immunoblot analysis of Campylobacter fetus lipopolysaccharides. 882 27

The Kinase-Splitting Membranal Proteinase (KSMP) is a metallo-endoproteinase that clips off the carboxyl terminus tail of the catalytic (C) subunit of protein kinase A to yield a truncated, catalytically inactive protein (C'). Here we report (a) a new procedure for the purification of KSMP, yielding a major protein band in SDS-polyacrylamide gel electrophoresis that correlates with the characteristic KSMP activity; (b) the sequence of tryptic peptides obtained from this band, suggesting an identity between this protein and meprin beta; (c) the immuno-detection by specific anti-peptide antibodies of both the alpha and the beta subunits of meprin in KSMP preparations; (d) the stable expression of meprin beta in a mammalian cell line (293) to establish a clone that constitutively expresses the full-length precursor of meprin beta; and (e) the optimalization of the proteolytic activation of this precursor to obtain an enzyme exhibiting the specific KSMP cleavage, suggesting that KSMP is either derived from, or identical with, the meprin beta gene product. It is hoped that these results will shed light on the possible physiological role of KSMP and the way it may affect protein kinase A-mediated processes.
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PMID:The cleavage of protein kinase A by the kinase-splitting membranal proteinase is reproduced by meprin beta. 893 81

Hepsin, a putative cell-surface serine proteinase, has been isolated from the microsomal membranes of rat liver and purified to homogeneity by hydroxyapatite, DEAE-Sepharose, and benzamidine-Sepharose chromatography. The course of purification was monitored using antibodies raised against a 20-mer peptide at the C-terminus of rat hepsin, and the identity of the purified protein was confirmed by partial amino-acid sequencing. A single-chain precursor of ca. 50 kDa found in the microsomes underwent spontaneous maturation in the course of purification so that the last, affinity chromatography, step recovered only the mature form which dissociated to subunits of 31 and 19 kDa under reducing SDS-PAGE. Proteinase digestion experiments with microsomal vesicles are consistent with the luminal orientation of the precursor C-terminus, which would result in its extracellular orientation upon transportation to the cell surface. [3H]diisopropylfluorophosphate covalently binds to the large subunit showing it to be the catalytic one. The N-terminal sequencing of this subunit demonstrates that the zymogen is converted to the active serine proteinase by cleavage at the Arg161-Ile162 site. Activity measurements with short synthetic peptides show that the enzyme cleaves after basic amino-acid residues, Arg being preferable to Lys. The inhibition pattern is typical of trypsin-like serine proteinases. The pH-dependence of activity within the range pH 6-9 has no maximum, the activity increasing continuously with pH. These results are consistent with the earlier predictions based on hepsin amino-acid sequence and elucidate the specificity and other earlier unknown enzymatic and molecular properties of the enzyme.
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PMID:Purification and characterization of hepsin from rat liver microsomes. 900 40

Pediocin PD-1, produced by Pediococcus damnosus NCFB 1832, is inhibitory to several food spoilage bacteria and food-borne pathogens. However, pediocin PD-1 is not active against other Pediococcus spp. and differs in this respect to other pediocins produced by Pediococcus acidilactici and Pediococcus pentosaceus. Production of pediocin PD-1 starts during early growth and reaches-a plateau at the end of exponential growth. Pediocin PD-1 was partially purified and its size was determined by tricine-SDS-PAGE as approximately 3.5 kDa. The isoelectric point (pI) of pediocin PD-1 is approximately 3.5, as determined with the Rotofor electrofocusing cell (BioRad). Pediocin PD-1 is heat-resistant (10 min at 121 degrees C) and remains active after 30 min of incubation at pH 2-10. Pediocin PD-1 is resistant to treatment with pepsin, papain, alpha-chemotrypsin and trypsin, but not Proteinase K. Pediocin PD-1 is bactericidal against sensitive cells of Oenococcus oeni (previously Leuconostoc oenos).
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PMID:Pediocin PD-1, a bactericidal antimicrobial peptide from Pediococcus damnosus NCFB 1832. 924 79

Antigens recognized by monoclonal antibodies (Mabs) raised to the surface of the obligate nematode hyperparasite Pasteuria penetrans were characterized. Using the attachment of spores of the bacterium to host nematodes to determine the biological variability present on the spore surface greatly underestimated the amount of surface heterogeneity present compared with estimates from immunological techniques. This heterogeneity differed not only between different individual spores from the same population but also between different spore populations. None of the Mabs completely inhibited any spore population from attaching to the nematode cuticle, suggesting that the mechanism of attachment may be more complex than previously supposed. Chemical degradation of one particular epitope recognized by monoclonal antibody PP1/117, and designated ep117, occurred after treatment with NaOH, periodate or Proteinase K, suggesting that an O-linked glycoprotein may be involved. Fibronectin, which had been found to bind to Pasteuria spores through hydrophobic interactions, also prohibited the Mab from recognizing ep117. However, SDS-PAGE of spore extracts followed by immunoblotting showed that none of the Mabs could detect this epitope and so ep117 may be conformational in nature. Thus, the conformation of any particular epitope recognized by a Mab may be important in determining to which nematode a particular spore will attach. The distribution of a particular epitope within a population of spores will in turn therefore determine its virulence on a particular nematode.
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PMID:Diversity and partial characterization of putative virulence determinants in Pasteuria penetrans, the hyperparasitic bacterium of root-knot nematodes (Meloidogyne spp.). 928 26

The implementation of convicted felon DNA databases by increasing numbers of forensic science laboratories has engendered the need for a quick, efficient, and cost-effective method for the isolation of DNA from liquid blood samples. Because of the large numbers of samples involved, the ideal method would combine high throughput capability with maximal yield, high quality, and minimal time. We have found that the QIAGEN QIAamp Blood Kit/Tissue Kit satisfy all of these requirements. This simple, low cost spin column procedure yields purified DNA of approximately 20-30 kb that can be used directly in PCR or other enzymatic reactions without further purification. We compared the QIAamp isolation procedure to the standard SDS-Proteinase K/organic extraction/microcon purification procedure currently used by many forensic laboratories. The QIAamp procedure consistently gave a two- to four-fold increased yield relative to the organic extraction procedure. The DNA obtained was of high molecular weight, exhibited little degradation, and was suitable for RFLP and PCR analyses. We have found QIAGEN's QIAamp DNA isolation procedure to be ideally suited for preparation of samples for DNA databasing.
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PMID:DNA extraction from liquid blood using QIAamp. 930 38


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