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The multicatalytic proteinase (prosome, proteasome): comparison of the eukaryotic and archaebacterial enzyme. 多催化蛋白酶(prosome, proteasome):真核生物和古细菌酶的比较。
Biomedica biochimica acta Pub Date : 1991-01-01
B Dahlmann, F Kopp, L Kuehn, R Hegerl, G Pfeifer, W Baumeister
{"title":"The multicatalytic proteinase (prosome, proteasome): comparison of the eukaryotic and archaebacterial enzyme.","authors":"B Dahlmann,&nbsp;F Kopp,&nbsp;L Kuehn,&nbsp;R Hegerl,&nbsp;G Pfeifer,&nbsp;W Baumeister","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Proteasomes isolated and purified from rat muscle tissue and from the archaebacterium Thermoplasma acidophilum have a very similar size and shape, but the subunit composition is less complex in the archaebacterium as compared to the eukaryotic particle. The archaebacterial enzyme contains a catalytic site with chymotryptic specificity, which is inhibited by serine proteinase inhibitors and clearly differs from the eukaryotic particle which has a minimum of three catalytic sites for peptide bond hydrolysis of a yet undefined mechanism.</p>","PeriodicalId":8948,"journal":{"name":"Biomedica biochimica acta","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1991-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12964700","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
The human cystatin gene family: cloning of three members and evolutionary relationship between cystatins and Bowman-Birk type proteinase inhibitors. 人胱抑素基因家族:三个成员的克隆及胱抑素与Bowman-Birk型蛋白酶抑制剂的进化关系。
Biomedica biochimica acta Pub Date : 1991-01-01
E Saitoh, S Isemura, K Sanada, K Ohnishi
{"title":"The human cystatin gene family: cloning of three members and evolutionary relationship between cystatins and Bowman-Birk type proteinase inhibitors.","authors":"E Saitoh,&nbsp;S Isemura,&nbsp;K Sanada,&nbsp;K Ohnishi","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Three genes from the human cystatin gene family have been isolated from a bacteriophage lambda library containing Hind III digests of human genomic DNA. The cloned genes were identified with three DNA probes each containing exon 1, exon 2 and exon 3 of the CST1 gene for cystatin SN. The genes, which we name CST2B, CST4, and CST5, are 6.8 kb, 5.4 kb and 12.5 kb in size, respectively. Statistical analysis of DNA sequence homology elucidated that the second and third exons of cystatin (family II) genes and three cystatin (family II) gene like segments in the kininogen (family III) genes are significantly homologous to the gene segments coding for the inhibitory domains of Bowman-Birk type proteinase inhibitors.</p>","PeriodicalId":8948,"journal":{"name":"Biomedica biochimica acta","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1991-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12965125","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Recombinant Q53E- and Q53N--chicken egg white cystatin variants inhibit papain, actinidin and cathepsin B. 重组Q53E-和Q53N-鸡蛋清胱抑素变体抑制木瓜蛋白酶、actitinidin和组织蛋白酶B。
Biomedica biochimica acta Pub Date : 1991-01-01
G Genenger, S Lenzen, R Mentele, I Assfalg-Machleidt, E A Auerswald
{"title":"Recombinant Q53E- and Q53N--chicken egg white cystatin variants inhibit papain, actinidin and cathepsin B.","authors":"G Genenger,&nbsp;S Lenzen,&nbsp;R Mentele,&nbsp;I Assfalg-Machleidt,&nbsp;E A Auerswald","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>A cloned synthetic gene coding for (AEF S1M M29I M89L) chicken egg white cystatin was modified site-specifically at position Q53 by cassette mutagenesis. Two recombinant variants were isolated from a pIN-III-ompA E. coli expression system and purified by Cm-papain affinity chromatography. The mutations at the position 53 were confirmed by amino acid composition and amino acid sequence analysis of the appropriate tryptic peptides. The complexes of both cystatin variants, the Q53E- and Q53N-variant with papain, display Ki values similar to those determined with native chicken cystatin. However, the Ki values of the complexes with actinidin are hundredfold and with cathepsin B three hundredfold higher than with the native chicken cystatin. The different inhibition kinetics of these variants compared to wild type chicken cystatin emphasizes the specificity of single amino acid substitutions for optimal contacts between the binding segments of enzyme and inhibitor.</p>","PeriodicalId":8948,"journal":{"name":"Biomedica biochimica acta","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1991-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12965128","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Effect of thermodynamic water activity on protease-catalyzed peptide synthesis in mainly organic media. 热力学水活度对主要有机介质中蛋白酶催化多肽合成的影响。
Biomedica biochimica acta Pub Date : 1991-01-01
P J Halling
{"title":"Effect of thermodynamic water activity on protease-catalyzed peptide synthesis in mainly organic media.","authors":"P J Halling","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The role of water in mainly organic reaction mixtures for biocatalysis is best analysed in terms of the thermodynamic water activity. This determines water mass action effects on the equilibria of protease-catalysed peptide or ester synthesis. It can also be useful to predict the amount of water bound by the enzyme, and hence its catalytic activity, as other factors are changed.</p>","PeriodicalId":8948,"journal":{"name":"Biomedica biochimica acta","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1991-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12981672","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
The synthesis of chromogenic peptide substrates containing p-nitroanilides of arginine and lysine, catalyzed by proteinases adsorbed on support material. 用吸附在载体上的蛋白酶催化合成含精氨酸和赖氨酸对硝基苯胺的显色肽底物。
Biomedica biochimica acta Pub Date : 1991-01-01
T L Voyushina, Terent'eva EYu, V M Stepanov
{"title":"The synthesis of chromogenic peptide substrates containing p-nitroanilides of arginine and lysine, catalyzed by proteinases adsorbed on support material.","authors":"T L Voyushina,&nbsp;Terent'eva EYu,&nbsp;V M Stepanov","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The synthesis of the chromogenic substrates for trypsin-like proteinases catalyzed by alpha-chymotrypsin and subtilisin from B. subtilis strain 72 were carried out in the organic media at a low water content using the enzymes adsorbed on different porous materials. The method proposed allows us to vary the structure of the compounds to be synthesized and is a suitable technique for their scaling-up.</p>","PeriodicalId":8948,"journal":{"name":"Biomedica biochimica acta","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1991-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12981720","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Use of Z-amino acid-glyceryl esters in protease catalyzed peptide synthesis. z -氨基酸-甘油酯在蛋白酶催化多肽合成中的应用。
Biomedica biochimica acta Pub Date : 1991-01-01
J Wiese, H G Gattner, H Zahn
{"title":"Use of Z-amino acid-glyceryl esters in protease catalyzed peptide synthesis.","authors":"J Wiese,&nbsp;H G Gattner,&nbsp;H Zahn","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The alpha-glyceryl esters of Z-Gly, Z-Phe and Z-Tyr were synthesized and their use for protease catalyzed peptide synthesis was studied. Three enzymes isolated from crude papain were compared in their catalytic potency. Syntheses with alpha-chymotrypsin were performed in a biphasic system.</p>","PeriodicalId":8948,"journal":{"name":"Biomedica biochimica acta","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1991-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12983040","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Preparative-scale enzyme-catalyzed peptide synthesis using solubilizing N-terminal protecting groups. 利用可溶n端保护基团的制备级酶催化肽合成。
Biomedica biochimica acta Pub Date : 1991-01-01
A Fischer, A Schwarz, C Wandrey, A S Bommarius, G Knaup, K Drauz
{"title":"Preparative-scale enzyme-catalyzed peptide synthesis using solubilizing N-terminal protecting groups.","authors":"A Fischer,&nbsp;A Schwarz,&nbsp;C Wandrey,&nbsp;A S Bommarius,&nbsp;G Knaup,&nbsp;K Drauz","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Several amino acid derivatives with the negatively charged N alpha-protecting groups Maleyl (Mal) and Citraconyl (Cit) were synthesized and used in enzyme-catalyzed peptide synthesis. Compared to commonly used alpha-amino protecting groups in chemical peptide synthesis (Z, Fmoc, Boc, etc.), these charged protecting groups strongly increase both water solubility of different aromatic amino acid derivatives and activities of synthesis reactions. As a consequence of the solubilizing effect, we used these groups in the kinetically controlled peptide synthesis choosing kyotorphin (tyrosyl-arginine) as a model peptide. With Mal-Tyr-OEt as substrate and Arg-OEt as nucleophile, we succeeded in the alpha-chymotrypsin-catalyzed production of about 12 kg of Tyr-Arg (50.4% overall yield) in a 300 l batch experiment.</p>","PeriodicalId":8948,"journal":{"name":"Biomedica biochimica acta","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1991-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"13001729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Erythrocyte rheology and lipid peroxidation in burns. 烧伤的红细胞流变学和脂质过氧化。
Biomedica biochimica acta Pub Date : 1991-01-01
G Bekyarova, T Yankova, D Yankov, I Kozarev
{"title":"Erythrocyte rheology and lipid peroxidation in burns.","authors":"G Bekyarova,&nbsp;T Yankova,&nbsp;D Yankov,&nbsp;I Kozarev","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Alterations in the erythrocyte rheology and the contents of activated free radical oxidation products (conjugated dienes, products of thiobarbituric acid and Schiff bases) in the acute phase of experimental thermic injury of the skin were studied. Erythrocyte flexibility reduction and erythrocyte aggregation increase correlated with elevated amounts of free radical oxidation products. Alpha-tocopherol avoided the accumulation of free radical oxidation products and improved both antioxidant defence and erythrocyte rheology. Thus we suppose that free radical oxidation products probably participate in the pathogenesis of erythrocyte rheology disturbances after thermic trauma.</p>","PeriodicalId":8948,"journal":{"name":"Biomedica biochimica acta","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1991-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"13020178","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Improved purification and characterization of membraneous and cytosolic inositol phospholipid-specific phospholipases C from porcine brain cortex. 改进了猪脑皮质膜质和细胞质肌醇磷脂特异性磷脂酶C的纯化和鉴定。
Biomedica biochimica acta Pub Date : 1991-01-01
B Hoffmann, C Seib, A Höer, D Höer, E Oberdisse, W Rosenthal, G Schultz
{"title":"Improved purification and characterization of membraneous and cytosolic inositol phospholipid-specific phospholipases C from porcine brain cortex.","authors":"B Hoffmann,&nbsp;C Seib,&nbsp;A Höer,&nbsp;D Höer,&nbsp;E Oberdisse,&nbsp;W Rosenthal,&nbsp;G Schultz","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>A phospholipase C was solubilized and purified from membranes of porcine brain cortex. Simultaneously, a phospholipase C was purified from a cytosolic fraction of porcine brain cortex. The enrichment of phospholipase C from either fraction was about 1000-fold as determined by hydrolysis of phosphatidylinositol 4,5-bisphosphate. Phospholipases C purified from membranes or from cytosol were indistinguishable with regard to the following properties: The enzyme activities copurified with a protein of 145 kDa. The standard sedimentation coefficients (s20,w values) of the purified enzymes were 6.2 S in the absence or presence of 0.3% (w/v) sodium cholate; Stokes' radii, estimated by gel filtration on a Superose 6 HR 10/30 column in the presence of 0.3% sodium cholate, were 4.5 nm; calculated molecular masses were about 120 kDa; no significant hydrolysis of phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine by preparations of purified phospholipase C was observed; adenine and guanine nucleotides affected the activity of purified enzymes in a complex manner. Thus, the enzymes purified from membraneous and from cytosolic fractions exhibited properties of the phospholipase C-beta form. The enzymes purified from either fraction required Ca2+ at a low concentration (100 nM to 10 microM) for maximal activity. The advantage of the present purification procedure is that the purified enzymes were free of phosphatidylinositol 4,5-bisphosphate 5-phosphatase, inositol 1,4,5-trisphosphate 5-phosphatase and guanine nucleotide-binding proteins after three chromatographic steps. The purified enzymes may, therefore, prove useful for studying the hormonal regulation of phospholipase C in reconstituted systems and for the preparation of [5-32P]inositol 1,4,5-trisphosphate from [5-32P]phosphatidylinositol 4,5-bisphosphate.</p>","PeriodicalId":8948,"journal":{"name":"Biomedica biochimica acta","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1991-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12816169","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Effect of ascorbic acid on hydroxyl radical generation by chemical, enzymatic and cellular systems. Importance for antioxidant prevention of pulmonary emphysema. 抗坏血酸对化学、酶和细胞系统生成羟基自由基的影响。抗氧化预防肺气肿的重要性。
Biomedica biochimica acta Pub Date : 1991-01-01
D Nowak, G Piasecka, A Antczak, T Pietras
{"title":"Effect of ascorbic acid on hydroxyl radical generation by chemical, enzymatic and cellular systems. Importance for antioxidant prevention of pulmonary emphysema.","authors":"D Nowak,&nbsp;G Piasecka,&nbsp;A Antczak,&nbsp;T Pietras","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The ability of ascorbic acid (AA) (25 to 500 microM) to increase OH production by a chemical (Fe(2+)-EDTA-H2O2), an enzymatic (xanthine-xanthine oxidase-Fe(2+)-EDTA) and a cellular system (3.10(6) human polymorphonuclear leukocytes (PMNL) or murine peritoneal macrophages (PM) activated with 7.2 ng PMA/ml) was studied. At all concentrations used AA strongly enhanced OH generation by the chemical and the enzymatic systems. However, the maximal increase of about 14-fold was found for incomplete chemical system (10 microM Fe(2+)-20 microM EDTA) and 500 microM AA. In the case of phorbol-myristate-acetate-activated-PMNL and macrophages, the moderate increase in OH formation was only caused by low AA concentrations. At 50 microM AA, the OH formation was 112 +/- 3 and 117 +/- 4% of control, respectively. Higher AA concentrations had no influence or even decreased OH formation by phagocytes. It is suggested that administration of AA will not significantly enhance OH generation from pulmonary phagocytes and could be useful for prevention of the oxidant-mediated lung injury related to inflammation.</p>","PeriodicalId":8948,"journal":{"name":"Biomedica biochimica acta","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1991-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12825023","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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