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酵母分泌突变体中外切葡聚糖酶活性的积累与分泌

Accumulation and secretion of exoglucanase activity in yeast secretory mutants.

作者信息

Hernández L M, Ramírez M, Olivero I, Larriba G

出版信息

Arch Microbiol. 1986 Dec;146(3):221-6. doi: 10.1007/BF00403220.

Abstract

Representative conditional yeast secretory mutants, blocked in transport of secretory and plasma membrane proteins from the endoplasmic reticulum (sec 18), from the Golgi body (sec 7) and in transport of secretory vesicles (sec 1), accumulated exoglucanase, a constitutive yeast activity, when incubated at the restrictive temperature (37 degrees C). Different proportions of the accumulated activity were released by mutant cells under permissive conditions. The presence or absence of cycloheximide during the secretion period made no differences in the results. More than 90% of the internal activity was bound to membrane in wild type cells. However, only the soluble pool underwent changes during the accumulation or secretion periods. The bulk of secretory invertase accumulated by sec 1 was also soluble. By contrast sec 7 and sec 18 accumulated membrane-bound as well as soluble invertase forms and both were secreted in similar proportions in each mutant. More than 90% of the accumulated invertase was secreted at the permissive temperature in sec 18 cells. That percentage was significantly lower for exoglucanase (less than 65%). Concomitantly, invertase accumulated by this mutant exited from the cells with a lower half time (t1/2 = 24 min) than accumulated exoglucanase (t1/2 = 150 min). These results may be interpreted assuming that exoglucanase is exported by a passive flow of the soluble pool.

摘要

典型的条件性酵母分泌突变体,在内质网(sec18)、高尔基体(sec7)分泌和质膜蛋白运输受阻以及分泌小泡运输受阻(sec1)时,当在限制温度(37摄氏度)下孵育时,会积累外切葡聚糖酶(一种组成型酵母活性)。在允许条件下,突变细胞释放出不同比例的积累活性。分泌期间有无放线菌酮对结果没有影响。在野生型细胞中,超过90%的内部活性与膜结合。然而,在积累或分泌期间,只有可溶性部分发生变化。sec1积累的大部分分泌型转化酶也是可溶的。相比之下,sec7和sec18积累了膜结合型和可溶性转化酶形式,并且在每个突变体中两者以相似的比例分泌。在允许温度下,sec18细胞中超过90%的积累转化酶被分泌。外切葡聚糖酶的这一百分比显著较低(低于65%)。与此同时,该突变体积累的转化酶从细胞中排出的半衰期(t1/2 = 24分钟)比积累的外切葡聚糖酶(t1/2 = 150分钟)短。这些结果可以解释为假设外切葡聚糖酶是通过可溶性部分的被动流动输出的。

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