HORMONE PROCESSING AND MEMBRANE-BOUND PROTEINASES IN YEAST
HORMONE PROCESSING AND MEMBRANE-BOUND PROTEINASES IN YEAST
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DOI：
10.1002/j.1460-2075.1985.tb02333.x
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发表时间：
1985-01-01
期刊：
影响因子：
11.4
通讯作者：
WOLF, DH
中科院分区：
文献类型：
--
作者：
ACHSTETTER, T;WOLF, DH
寻找交配激素（信息素）α的前体蛋白的成熟肽酶使用代表前体蛋白的那些序列的短模型肽来进行酿酒酵母的因子的切割，其中切割被认为在体内发生。这项研究是在缺乏几种非特异性液泡肽酶的突变体中进行的。显色肽Cbz-Tyr-Lys-Arg-4-nitroanilide导致检测到一种称为蛋白酶yscF的膜结合酶。合成肽衍生物的切割发生在碱性氨基酸对之后，这是激素加工的建议信号。反应的最适pH为7.2。该酶在单个碱性氨基酸残基后不裂解，表明其不同于胰蛋白酶样蛋白酶。Triton X-100可增强蛋白水解活性。该酶能被EGTA、EDTA和汞剂强烈抑制，但对苯甲基磺酰氟不敏感。该酶的活性强烈依赖于Ca ~（2+）离子。在突变体（kex 2）中，其积累过糖基化的α-葡萄糖基化酶。膜结合的肽酶活性可能涉及去除保留在α-因子前体羧基末端的乙酰基和赖氨酰残基。通过使用模型肽Cbz-Tyr-Lys-Arg和Cbz-Tyr-Lys，可以鉴定前体分子初始切割后的因子信息素肽。
A search for maturating peptidases of the precursor protein of the mating hormone (pheromone) .alpha.-factor of Saccharomyces cerevisiae was performed using short model peptides representing those sequences of the precursor protein, where cleavage is thought to occur in vivo. This search was done in a mutant lacking several of the unspecific vacuolar peptidases. The chromogenic peptide Cbz-Tyr-Lys-Arg-4-nitroanilide led to the detection of a membrane-bound enzyme called proteinase yscF. Cleavage of the synthetic peptide derivative occurs after the basic amino acid pair, a proposed signal for hormone processing. Optimum pH for the reaction is 7.2. The enzyme does not cleave after single basic amino acid residues, indicating that it is distinct from trypsin-like proteinases. Proteolytic activity is enhanced by Triton X-100. The enzyme is strongly inhibited by EGTA, EDTA and mercurials but insensitive to phenylmethylsulfonyl fluoride. The enzyme activity is strongly dependent on Ca2+ ions. In a mutant (kex2), which accumulates an over-glycosylated .alpha.-factor precursor, no proteinase yscF activity can be found. Membrane-bound peptidase activity possibly involved in removal of the arginyl and lysyl residues remaining at the carboxy terminus of the .alpha.-factor pheromone peptide after the initial cut of the precursor molecule could be identified by using the model peptides Cbz-Tyr-Lys-Arg and Cbz-Tyr-Lys.