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)

Rat liver esterases focusing at pH 5.0 (referred to below as pI-5.0 esterases) are structurally related glycoproteins which differ slightly in their mobility in sodium dodecyl sulphate/polyacrylamide gel electrophoresis (SDS-PAGE). They reside in the lumen of the endoplasmic reticulum. We have studied their biosynthesis in cell-free systems programmed by total liver RNA, using sheep and rabbit antibodies to isolate the translation products related to these enzymes. Our results show that they are assembled as a precursor polypeptide chain (62 kDa) larger than the mature proteins. The pI-5.0 esterase mRNA could be extracted from bound but not free polysomes. Reticulocyte lysates supplemented with dog pancreas microsomes produced four esterase-related components in segregated form (61, 60, 58 and 56 kDa). The largest three correspond in electrophoretic mobility to the mature enzymes. They are glycoproteins that bind to concanavalin A, and can be reduced to the size of the shortest component by endo-beta-N-acetylglucosaminidase H (endo-H). Immunoprecipitation after biosynthetic labeling of the proteins in cultured hepatocytes also gave three glycosylated components that had the same mobility in SDS-PAGE as the mature enzymes. When tunicamycin was present in the culture medium, a single immunoprecipitable form was observed. Its apparent Mr was similar to that of the unglycosylated pI 5.0 esterase form synthesized in vitro in the presence of dog pancreas microsomes. Thus the biosynthesis of these esterases has characteristics in common with that of numerous secretory proteins, except for the rather large difference in size (approximately equal to 6 kDa) resulting from the proteolytic processing of their in-vitro-synthesized precursor.
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PMID:Biosynthesis of rat-liver pI-5.0 esterases in cell-free systems and in cultured hepatocytes. 308 77

An enzyme, esterase A1, which hydrolyzes tosyl-arginine methyl ester (Tos-Arg-OMe) was separated from esterase A2 and kallikrein of male rat urine and purified by a procedure involving ammonium sulfate fractionation, ion exchange chromatography, hydrophobic chromatography and gel filtration. The resulting preparation was apparently homogeneous, as assessed by polyacrylamide gel electrophoresis. The molecular weight of the preparation was estimated to be 27,000 by SDS-polyacrylamide gel electrophoresis and 30,000 by gel filtration. The enzyme was more specific for arginine methyl esters than for lysine methyl esters. The optimum pH determined with Tos-Arg-OMe as a substrate was 8.0 and the Km was 11.8 mM. The Tos-Arg-OMe esterolytic activity of esterase A1 was inhibited by soybean trypsin inhibitor, but not by aprotinin. In immunodiffusion analysis, the antiserum to esterase A1 formed immunoprecipitin arcs with this enzyme and the urine collected from rat bladder, but not with esterase A2, kallikrein, plasma and the urine collected from ureters. These results indicate that rat urinary esterase A1 differs from esterase A2 and kallikrein. The esterase A1 appears to be produced by accessory sex glands and excreted via the spermiduct into the urine.
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PMID:Purification and characterization of rat urinary esterase A1. 309 89

A heat-stable esterase has been purified 1080-fold to electrophoretic homogeneity from Sulfolobus acidocaldarius, a thermoacidophilic archaebacterium; 20% of the starting activity is recovered. The purified enzyme shows a specific activity of 158 units/mg, based on the hydrolysis of p-nitrophenyl acetate. The esterase hydrolyses short-chain p-nitrophenyl esters, aliphatic esters and triacylglycerols. It is strongly inhibited by paraoxon and phenylmethanesulphonyl fluoride, but only weakly by eserine. From sedimentation-equilibrium data and molecular sieving in polyacrylamide gels, the Mr of the esterase is estimated to be 117000-128000. SDS/polyacrylamide-gel electrophoresis reveals a single band of protein, of Mr 32000. The purified esterase crystallizes in the presence of poly(ethylene glycol) in short rods. The enzyme is inactivated only on prolonged storage at temperature above 90 degrees C.
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PMID:Purification and characterization of a heat-stable esterase from the thermoacidophilic archaebacterium Sulfolobus acidocaldarius. 312 84

Both the male and the female of Mastomys natalensis, an African rat, have high levels of nerve growth factor (NGF) in their submaxillary glands. Mastomys high molecular weight (HMW) NGF was purified by gel filtration and ion-exchange chromatography and was compared with HMW NGF from the male mouse submaxillary gland. Mastomys HMW NGF sediments as a 5S species, does not exhibit esterase activity, and is more difficult to dissociate at acid pH than mouse 7S NGF. The biological activity could be isolated as a purified Mastomys beta NGF protein identical in size and charge with that purified from male mice. The N-terminal amino acid sequence of the first 20 residues was determined and found to differ from that of mouse only at residue 8. Western blotting of Mastomys 5S NGF using antiserum against mouse beta NGF indicates that the beta NGF subunit of Mastomys is very similar to that of the mouse. Southern blots using a mouse kallikrein probe also demonstrate the presence of a large kallikrein family in Mastomys similar to that in mouse, and Northern blots verify transcription of kallikreins in Mastomys submaxillary gland. SDS-PAGE and isoelectric focusing gels reveal a Mastomys subunit that comigrates with mouse alpha subunit. However, neither oligonucleotide probes directed against mouse alpha subunit RNA nor antibodies directed against mouse alpha NGF cross-react strongly with the Mastomys material. This indicates that the second subunit of the Mastomys complex is not very similar to the mouse alpha subunit.
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PMID:The high molecular weight nerve growth factor complex from Mastomys natalensis differs from the murine nerve growth factor complex. 319 80

Human peripheral blood mononuclear cells, activated for 14 to 20 days with 1000 U/ml rIL-2, develop strong cytotoxicity for NK sensitive and resistant targets. This process is accompanied by the acquisition of cytoplasmic granules in approximately 60% of the cells and by the expression of esterase activity cleaving the synthetic substrate BLT. The esterase activity, localized in the cytoplasmic granules, was purified and characterized. Three proteins with 3H-DFP binding activity were isolated and had the following properties. Following the proposed nomenclature by Masson et al., the esterases were named human granzymes 1, 2, and 3. Human granzyme 1 on SDS-PAGE has an unreduced relative m.w. of 43,000 and can form disulfide-linked oligomers of relative higher m.w. All forms of granzyme 1 bind 3H-DFP. Upon reduction, granzyme 1 migrates with Mr 30,000 on SDS-PAGE. Additional proteolytic fragments of Mr 24,000 and Mr 28,000 are observed in some reduced preparations. Granzyme 1 cleaves the substrate BLT and appears homologous with murine granzyme A. Human granzyme 2 has an unreduced relative m.w. of 30,000; after reduction, it migrates at Mr 32,000. Even though granzyme 2 binds 3H-DFT, it does not cleave BLT. Human granzyme 2 has properties similar to those of murine granzymes B-H. Human granzyme 3 has unreduced and reduced relative m.w. of 25,000 and 28,000, respectively. It is active in cleaving the substrate BLT. A murine analog for human granzyme 3 has not been described previously. N-terminal sequencing of the purified human granzymes revealed that human granzyme 1 is the gene product of human Hanuka factor cDNA clone and that it represents the human homolog to murine granzyme A. Similarly, human granzyme 2 revealed absolute identity with cDNA-derived N-terminal sequence of a putative human lymphocyte protease cDNA clone.
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PMID:Characterization of three serine esterases isolated from human IL-2 activated killer cells. 326 82

A thrombin-like enzyme (I-SII-R) from the venom of the Brazilian snake Bothrops insularis (jararaca ilhoa) was purified to homogeneity by gel filtration on Sephadex G-150 followed by precipitation of contaminating proteins with half-saturated NaCl and two further gel filtrations under the same conditions. I-SII-R showed specific activities of 820 clotting units/mg protein and 530 TAME units/mg protein (a 23-fold purification) and a single precipitation band against antibothropic horse antiserum by immunoelectrophoresis, as well as a single band by PAGE and by SDS-PAGE with beta-mercaptoethanol. The estimated molecular weight was 45,000, of which the neutral sugars account for 9900, or 22%. Its amino acid composition, which accounts for a minimum mol. wt of 33,900 is as follows: Asx35, Thr18, Ser17, Glx15, Pro29, Gly22, Ala27, Val21, Cys18, Met7, Ile19, Leu25, Tyr11, Phe11, His7, Lys8, Arg17, Trp5. When compared to thrombin, I-SII-R is relatively stable at 45 degrees C and 60 degrees C, pH 7.4, but is inhibited by heparin at a higher rate than thrombin. Both enzymes hydrolyze the alpha and beta chains of fibrinogen and lose more than 95% of their clotting and esterase activities when treated with phenylmethylsulphonyl fluoride. In contrast to thrombin, I-SII-R does not activate factor XIII.
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PMID:Isolation and characterization of a thrombin-like enzyme from the venom of the snake Bothrops insularis (jararaca ilhoa). 343 90

Biosynthesis of the rat liver microsomal esterase with pI 6.1 was investigated in cell-free systems and in cultured hepatocytes, by using a rabbit antiserum. Protein synthesis directed by total rat liver RNA in wheatgerm extract or reticulocyte lysate generated a single immunoprecipitable product, also found with the RNA extracted from bound, but not from free, polysomes. When dog pancreas microsomal fractions were included, reticulocyte lysates gave two processed products, a prominent one slightly larger, and another slightly smaller, than the precursor, both resistant to exogenous proteinases and, hence, segregated within vesicles. The processing was co-translational; it consisted of the removal of a peptide fragment and, for the large component, the addition of a single oligosaccharide chain. Indeed, this component bound to concanavalin A-Sepharose and gave the small one (approximately 2000 Mr loss) by cleavage with endo-beta-N-acetylglucosaminidase H (endo-H). A single labelled peptide was precipitated from hepatocytes incubated with [35S]methionine. Its apparent Mr was decreased by approximately 2000 after treatment with endo-H; it was then identical with that of an unglycosylated form produced in hepatocytes poisoned with tunicamycin. Even in that case, immunoreactive peptides were not detected in the culture medium. Whether synthesized in reticulocyte lysate or in hepatocytes, the glycosylated forms migrated in SDS/polyacrylamide-gel electrophoresis as the purified enzyme labelled with [3H]di-isopropyl fluorophosphate. Thus, although pI-6.1 esterase is not secreted, its biosynthesis is, as yet, indistinguishable from that of secretory proteins. Its oligosaccharide moiety is apparently not the structural element that retains it in the endoplasmic reticulum.
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PMID:Biosynthesis of rat liver pI-6.1 esterase, a carboxylesterase of the cisternal space of the endoplasmic reticulum. 343 65

We have determined the nucleotide sequence of the ereB gene of plasmid pIP1527 which confers high-level resistance to erythromycin by inactivation in Escherichia coli. The open reading frame of the ereB gene, 1257-bp, was defined by initiation and termination codons and by cloning in vitro. The corresponding protein has a calculated Mr of 48,118 in close agreement with a previous estimation, 51,000, by electrophoresis of minicell extracts in SDS-polyacrylamide gels. The structure of the modified erythromycin was determined by physico-chemical techniques including mass spectrometry, infrared spectrophotometry and 13C nuclear magnetic resonance. The data obtained indicated that like ereA (Ounissi and Courvalin, 1985) ereB encodes an erythromycin esterase. Comparison of the amino acid sequences of the two isozymes did not reveal any statistically significant homology. Analysis of the nucleotide sequence of the ereB gene suggests that this resistance determinant should be exogenous to E. coli.
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PMID:Analysis of the nucleotide sequence of the ereB gene encoding the erythromycin esterase type II. 352 38

The thiol-dependent serine proteinase (inhibited by DFP, PMSF, pCMB and iodoacetate) was isolated from the whole krill specimens and from the content of the krill digestive tract. The enzyme was purified to homogeneity using a seven-step procedure. Its specific activity with denatured haemoglobin as a substrate was about 6.0 unit/mg. The molecular weight of the enzyme, as determined by gel exclusion chromatography was 33 000 and by polyacrylamide gel electrophoresis with SDS 31 600 (12.5% gel) and 27 000 (7.5% gel). The enzyme is an acidic glycoprotein (pI below 2.9) containing about 5% of carbohydrate. The pH optimum of the enzyme with haemoglobin was 6.0 at the optimal temperature of 40 degrees C in 15-min reaction. The enzyme showed the esterase activity (hydrolysis of BAEE) and was inactive with carbobenzoxy- and benzoyl-dipeptides with the following C-terminal amino acids: Phe, Tyr, Lys, Gly and Leu.
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PMID:Purification and characterization of a proteinase from Euphausia superba Dana (Antarctic krill). 353 51

The identity of a peptidase activity with human serum pseudocholinesterase (PsChE) purified to apparent homogeneity was demonstrated by co-elution of both peptidase and PsChE activities from procainamide-Sepharose and concanavalin-A--Sepharose affinity chromatographic columns; comigration on polyacrylamide gel electrophoresis; co-elution on Sephadex G-200 gel filtration and coprecipitation at different dilutions of an antibody raised against purified PsChE. The purified enzyme showed a single protein band on gel electrophoresis under non-denaturing conditions. SDS gel electrophoresis under reducing conditions, followed by silver staining, also gave a single protein band (Mr approximately equal to 90,000). Peptidase activity using different peptides showed the release of C-terminal amino acids. Blocking the carboxy terminal by an amide or ester group did not prevent the hydrolysis of peptides. There was no evidence for release of N-terminal amino acids. Potent anionic or esterase site inhibitors of PsChE, such as eserine sulphate, neostigmine, procainamide, ethopropazine, imipramine, diisopropylfluorophosphate, tetra-isopropylpyrophosphoramide and phenyl boronic acid, did not inhibit the peptidase activity. An anionic site inhibitor (neostigmine or eserine) in combination with an esterase site inhibitor (diisopropylfluorophosphate) also did not inhibit the peptidase. However, the choline esters (acetylcholine, butyrylcholine, propionylcholine, benzoylcholine and succinylcholine) markedly inhibited the peptidase activity in parallel to PsChE. Choline alone or in combination with acetate, butyrate, propionate, benzoate or succinate did not significantly inhibit the peptidase activity. It appeared that inhibitor compounds which bind to both the anionic and esteratic sites simultaneously (like the substrate analogues choline esters) could inhibit the peptidase activity possibly through conformational changes affecting a peptidase domain.
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PMID:A peptidase activity exhibited by human serum pseudocholinesterase. 354 20


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