首页> 外文期刊>Cell biology international. >Single amino acid mutations in the Saccharomyces cerevisiae Saccharomyces cerevisiae rhomboid peptidase, Pcp1p, alter mitochondrial morphology
【24h】

Single amino acid mutations in the Saccharomyces cerevisiae Saccharomyces cerevisiae rhomboid peptidase, Pcp1p, alter mitochondrial morphology

机译:单氨基酸突变在酿酒酵母酿酒酵母酿酒酵母霉菌肽肽酶,PCP1P,改变线粒体形态

获取原文
获取原文并翻译 | 示例
           

摘要

Abstract Key to mitochondrial activities is the maintenance of mitochondrial morphology, specifically cristae structures formed by the invagination of the inner membrane that are enriched in proteins of the electron transport chain. In Saccharomyces cerevisiae , these cristae folds are a result of the membrane fusion activities of Mgm1p and the membrane‐bending properties of adenosine triphosphate (ATP) synthase oligomerization. An additional protein linked to mitochondrial morphology is Pcp1p, a serine protease responsible for the proteolytic processing of Mgm1p. Here, we have used hydroxylamine‐based random mutagenesis to identify amino acids important for Pcp1p peptidase activity. Using this approach we have isolated five single amino acid mutants that exhibit respiratory growth defects that correlate with loss of mitochondrial genome stability. Reduced Pcp1p protease activity was confirmed by immunoblotting with the accumulation of improperly processed Mgm1p. Ultra‐structural analysis of mitochondrial morphology in these mutants found a varying degree of defects in cristae organization. However, not all of the mutants presented with decreased ATP synthase complex assembly as determined by blue native polyacrylamide gel electrophoresis. Together, these data suggest that there is a threshold level of processed Mgm1p required to maintain ATP synthase super‐complex assembly and mitochondrial cristae organization.
机译:摘要线粒体活动的关键是维持线粒体形态,特别是由富含电子传递链蛋白质的内膜内陷形成的嵴结构。在酿酒酵母中,这些嵴折叠是Mgm1p的膜融合活性和三磷酸腺苷(ATP)合酶寡聚的膜弯曲特性的结果。另一种与线粒体形态相关的蛋白质是Pcp1p,这是一种丝氨酸蛋白酶,负责Mgm1p的蛋白水解过程。在这里,我们使用基于羟胺的随机突变来鉴定对Pcp1p肽酶活性重要的氨基酸。利用这种方法,我们分离出了五种单氨基酸突变体,它们表现出与线粒体基因组稳定性丧失相关的呼吸生长缺陷。Pcp1p蛋白酶活性降低通过免疫印迹法证实,Mgm1p处理不当会导致Pcp1p蛋白酶活性降低。对这些突变体线粒体形态的超微结构分析发现嵴组织存在不同程度的缺陷。然而,根据蓝色天然聚丙烯酰胺凝胶电泳测定,并非所有突变体的ATP合成酶复合物组装都减少。总之,这些数据表明,维持ATP合成酶超复杂组装和线粒体嵴组织所需的加工Mgm1p有一个阈值水平。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号