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首页> 外文期刊>Molecular and Cellular Biology >Characterization of the yeast KEX1 gene product: a carboxypeptidase involved in processing secreted precursor proteins.
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Characterization of the yeast KEX1 gene product: a carboxypeptidase involved in processing secreted precursor proteins.

机译：酵母KEX1基因产物的表征：一种参与加工分泌的前体蛋白的羧肽酶。

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摘要

We have identified and partially characterized the Saccharomyces cerevisiae KEX1 gene product, Kex1p, to assess its role in processing secreted protein precursors. Anti-Kex1p antibodies identified a 113-kilodalton protein that was absent in cells in which the KEX1 gene has been disrupted and that was more abundant in cells overexpressing the KEX1 gene. Kex1p was found to be a membrane-associated glycoprotein with N-linked carbohydrate. The N-linked oligosaccharide(s) was modified in a progressive manner after synthesis, causing the glycoprotein to slowly increase in mass to 115 kilodaltons. After a Kex2p-mediated cleavage event at specific pairs of basic amino acids, alpha-factor and K1 killer toxin precursors have COOH-terminal dibasic residue extensions and require a carboxypeptidase B-like enzyme to process the precursors to maturity. A carboxypeptidase activity, with apparent specificity for basic amino acids, was detected in KEX1 cells. Disruption of the KEX1 gene abolished this activity, while overexpression of KEX1 increased it. Our results provide biochemical evidence consistent with earlier genetic work, that KEX1 encodes a serine carboxypeptidase involved in the processing of precursors to secreted mature proteins.
机译：我们已经鉴定并部分表征了酿酒酵母KEX1基因产物Kex1p，以评估其在加工分泌的蛋白质前体中的作用。抗Kex1p抗体鉴定出一种113公斤的蛋白质，该蛋白质在KEX1基因已被破坏的细胞中不存在，而在过表达KEX1基因的细胞中更为丰富。发现Kex1p是具有N-连锁碳水化合物的膜相关糖蛋白。 N-连接的寡糖在合成后以渐进方式进行修饰，使糖蛋白的质量缓慢增加至115千道尔顿。在特定的碱性氨基酸对上发生Kex2p介导的裂解事件后，α因子和K1杀伤毒素前体具有COOH末端的二元残基延伸，并需要羧肽酶B样酶来处理前体至成熟。在KEX1细胞中检测到了对碱性氨基酸具有明显特异性的羧肽酶活性。 KEX1基因的破坏取消了该活性，而KEX1的过表达则增加了该活性。我们的结果提供了与早期遗传工作一致的生化证据，即KEX1编码丝氨酸羧肽酶，参与分泌成熟蛋白的前体加工。

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