Limited proteolysis of high molecular weight histidine-rich protein of rat epidermis by epidermal proteinases.
Limited proteolysis of high molecular weight histidine-rich protein of rat epidermis by epidermal proteinases.
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表皮蛋白酶对大鼠表皮高分子量富含组氨酸的蛋白质进行有限的蛋白水解。
DOI：
10.1111/1523-1747.ep12462067
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Epstein,WL
中科院分区：
文献类型：
--
作者：
Kashima,M;Fukuyama,K;Kikuchi,M;Epstein,WL
通过SDS-PAGE分析，研究了可能参与表皮富含组氨酸蛋白Mr> 300k蛋白水解的表皮蛋白酶。在柠檬酸-蔗糖溶液中，从2日龄大鼠颗粒细胞中提取富含组氨酸的Mr> 300k蛋白，通过sepphacryl S-300柱层析与蛋白酶和较小的Mr蛋白分离。无蛋白酶富组氨酸蛋白在pH 3.5-9、37℃条件下稳定12 h。从大鼠表皮中部分纯化蛋白酶，并测定每种酶的抑制剂谱。对富含组氨酸的Mr56k蛋白进行有限的水解，得到了含有组织蛋白酶D的Mr56k衍生物，pH值为3.5-7.5，丝氨酸蛋白酶的pH值为7-9。在pH为3.5和7.5的条件下，组织蛋白酶D还检测到Mr56k蛋白进一步水解为Mr44k和Mr45k和47k的双链，而丝氨酸蛋白酶将Mr56k蛋白降解为多个蛋白带。组织蛋白酶B和L可降解Mr bbb30k蛋白，但未发现特异性优势产物。我们认为组织蛋白酶D和丝氨酸蛋白酶可能在玉米凝固过程中对富含组氨酸的蛋白质的原位加工中发挥作用。
Epidermal proteinases, which may be involved in proteolysis of Mr> 300k histidine-rich protein in epidermis, were studied by SDS-PAGE analysis. Mr> 300k histidine-rich protein was extracted from granular cells of 2-day-old rats in citric acid-sucrose solution and separated from proteinases and smaller Mrproteins by Sephacryl S-300 column chromatography. The proteinase-free histidine-rich protein was stable in pH 3.5-9 at 37°C for 12 h. Proteinases were partially purified from rat epidermis and inhibitor spectrum determined for each enzyme. Limited hydrolysis of Mr> 300k histidine-rich protein yielded a derivative of Mr56k with cathepsin D at pH 3.5-7.5 and a serine proteinase at pH 7-9. Further proteolysis of Mr56k protein to Mr44k and a doublet of Mr45k and 47k also was detected with cathepsin D at pH 3.5 and 7.5, respectively, while the serine proteinase degraded Mr56k protein to a number of protein bands. Cathepsins B and L degraded Mr> 300k protein but no specific predominant product was identified. We suggest that cathepsin D and the serine proteinase may play a role in in situ processing of histidine-rich protein during cornification.