PROTEASE ACTIVITY IN BROOK TROUT (SALVELINUS-FONTINALIS) FOLLICLE WALLS DEMONSTRATED BY SUBSTRATE-POLYACRYLAMIDE GEL-ELECTROPHORESIS
PROTEASE ACTIVITY IN BROOK TROUT (SALVELINUS-FONTINALIS) FOLLICLE WALLS DEMONSTRATED BY SUBSTRATE-POLYACRYLAMIDE GEL-ELECTROPHORESIS
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DOI：
10.1095/biolreprod38.2.511
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发表时间：
1988-03-01
影响因子：
3.6
通讯作者：
GOETZ, FW
中科院分区：
文献类型：
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作者：
BERNDTSON, AK;GOETZ, FW
通过将蛋白质底物掺入十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（底物-SDS-PAGE）的测定系统，在溪鳟（Salvelinus fontinalis）卵母细胞周围的卵泡壁中证明了蛋白水解活性。当毛囊壁提取物在含有明胶的凝胶中进行电泳和孵育时，至少存在六种蛋白水解酶（78、70、67、59、22 和 20 kDa）。在这六种酶中，当毛囊壁提取物在含有酪蛋白的凝胶中溶解和孵育时，仅存在两种酶（20 和 22 kDa）。 78 和 70 kDa 酶的活性被金属蛋白酶抑制剂和溶胶原蛋白酶抑制剂完全抑制，并被丝氨酸蛋白酶抑制剂部分抑制。 67 和 59 kDa 酶的活性被金属蛋白酶抑制剂和胶原蛋白酶抑制剂完全阻断。丝氨酸蛋白酶抑制剂仅略微降低 22 和 20 kDa 酶的活性。
Proteolytic activity was demonstrated in the follicle wall surrounding oocytes of brook trout (Salvelinus fontinalis) by an assay system that incorporated protein substrates into sodium dodecyl sulfate-polyacrylamide gel electrophoresis (substrate-SDS-PAGE). At least six proteolytic enzymes (78, 70, 67, 59, 22 and 20 kDa) were present when follicle wall extracts were electrophoresed and incubated in gels containing gelatin. Of these six enzymes, only two enzymes (20 and 22 kDa) were present when follicle wall extracts were resolved and incubated in gels containing casein. The activities of the 78 and 70 kDa enzymes were completely inhibited with metallo- and collagenolytic protease inhibitors and partially inhibited with serine protease inhibitors. The activities of the 67 and 59 kDa enzymes were completely blocked with metallo- and collagenolytic protease inhibitors. The activities of the 22 and 20 kDa enzymes were only slightly decreased with a serine protease inhibitor.