CHARACTERIZATION OF THE INDIVIDUAL COLLAGENASES FROM CLOSTRIDIUM-HISTOLYTICUM
CHARACTERIZATION OF THE INDIVIDUAL COLLAGENASES FROM CLOSTRIDIUM-HISTOLYTICUM
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DOI：
10.1021/bi00308a036
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发表时间：
1984-01-01
期刊：
影响因子：
2.9
通讯作者：
VANWART, HE
中科院分区：
文献类型：
--
作者：
BOND, MD;VANWART, HE
六种胶原蛋白。度量。,可以利用。，。delta，。和.zeta.)是先前从溶组织梭菌中分离得到的，并对其进行了详细的鉴定。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测定的分子量范围为68,000-125,000。等电聚焦实验表明，胶原酶的等电点在5.35 ~ 6.20范围内。α的亚种。-和。gamma。-collagenases (.alpha。1 vs.。2和。1 vs.。2)等电点不同，但MW相同。微观不均一性也被观察到。-和。epsilon -胶原酶。测定了所有6种胶原酶的氨基酸组成，对中性糖和己糖的分析表明，这些酶都没有显著的碳水化合物含量。锌和钙是唯一能与胶原酶结合的金属。纯化的酶含有。apprx。1 mol锌/mol蛋白质，Ca含量从。是2 mol/mol。-胶原酶。-胶原酶为7mol /mol。所有的胶原酶对明胶的活性都是胶原蛋白的5-10倍。.alpha。度量。-和。gamma。-胶原酶对合成肽底物的活性明显低于。delta -，。epsilon。-和-胶原酶。溶组织菌产生2种不同的胶原酶。
Six collagenases (.alpha., .beta., .gamma., .delta., .epsilon. and .zeta.), previously isolated from C. histolyticum, were characterized in detail. The MW determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis range from 68,000-125,000. Isoelectric focusing experiments demonstrate that the isoelectric points of the collagenases are in the 5.35-6.20 range. The subspecies of .alpha.- and .gamma.-collagenases (.alpha.1 vs. .alpha.2 and .gamma.1 vs. .gamma.2) have different isoelectric points but the same MW. Microheterogeneity was also observed for the .beta.- and .epsilon.-collagenases. The amino acid compositions of all 6 collagenases were determined, and analysis for neutral sugars and hexosamines shows that none of the enzymes have a significant carbohydrate content. Zn and Ca are the only metals that copurify with the collagenases. The purified enzymes contain .apprx. 1 mol of Zn/mol of protein and a Ca content that varies from .apprx. 2 mol/mol for .alpha.-collagenase to .apprx. 7 mol/mol for .beta.-collagenase. All of the collagenases are 5-10 times more active against gelatin than collagen. The .alpha.-, .beta.- and .gamma.-collagenases are significantly less active toward the synthetic peptide substrates examined than the .delta.-, .epsilon.- and .zeta.-collagenases. C. histolyticum produces 2 distinct classes of collagenase.