CHARACTERIZATION OF CATHEPSIN-B AND COLLAGENOLYTIC CATHEPSIN FROM HUMAN PLACENTA
CHARACTERIZATION OF CATHEPSIN-B AND COLLAGENOLYTIC CATHEPSIN FROM HUMAN PLACENTA
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DOI：
10.1111/j.1432-1033.1978.tb12071.x
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发表时间：
1978-01-01
期刊：
影响因子：
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通讯作者：
ETHERINGTON, DJ
中科院分区：
文献类型：
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作者：
EVANS, P;ETHERINGTON, DJ
人胎盘组织蛋白酶B [EC 3.4.22.1]和胶原溶解组织蛋白酶[EC 3.4.22.-]在Amberlite CG-50柱上通过色谱分离。用DEAE-Sephadex（A-50）和Sephadex G-100柱层析法部分纯化了溶胶原组织蛋白酶。组织蛋白酶B通过CM-纤维素和Sephadex G-100层析纯化。这两种酶都需要通过硫醇化合物活化，并与有机汞-琼脂糖-4B结合。巯基封闭剂是抑制性的，这证实了一个必不可少的巯基存在。这些酶在pH3.5和28 ℃下降解端肽区的可溶性小牛皮肤胶原和不溶性牛腱胶原。C主要产生α-链条组件发现溶胶原组织蛋白酶不水解任何测试的低分子量合成底物。亮抑酶肽，一种结构类似物的甜菜碱含合成底物，和antipain，木瓜蛋白酶的抑制剂，强烈抑制这两种酶。两种酶的等电点相似，组织蛋白酶B为5.4，溶胶原组织蛋白酶为5.1。从Sephadex G-100上的层析，计算出组织蛋白酶B的MW为24，500，胶原溶解性组织蛋白酶的MW为34，600。
Human placental cathepsin B [EC 3.4.22.1] and collagenolytic cathepsin [EC 3.4.22.-] were separated by chromatography on columns of Amberlite CG-50. Collagenolytic cathepsin was partially purified by chromatography on DEAE-Sephadex (A-50) and Sephadex G-100. Cathepsin B was purified by chromatography on CM-cellulose and Sephadex G-100. Both enzymes required activation by thiol compounds and were bound to organomercurial-Sepharose-4B. Sulfhydryl-blocking reagents were inhibitory, which confirmed an essential thiol group to be present. The enzymes degraded soluble calf skin collagen and insoluble bovine tendon collagen in the telopeptide region at pH 3.5 and 28.degree. C to yield mainly .alpha.-chain components. Collagenolytic cathepsin was found not to hydrolyse any of the low-MW synthetic substrates that were tested. Leupeptin, a structural analog of arginine-containing synthetic substrates, and antipain, an inhibitor of papain, were strongly inhibitory to both enzymes. The isoelectric points of the enzymes were similar, being 5.4 for cathepsin B and 5.1 for collagenolytic cathepsin. From chromatography on Sephadex G-100 the MW of cathepsin B was calculated to be 24,500 and that of collagenolytic cathepsin to be 34,600.