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粗糙脉孢菌复合体I 17.8 kDa亚基的克隆、体外线粒体导入及膜组装

Cloning, in vitro mitochondrial import and membrane assembly of the 17.8 kDa subunit of complex I from Neurospora crassa.

作者信息

Azevedo J E, Abrolat-Scharff J, Eckerskorn C, Werner S

机构信息

Institut für Physiologische Chemie, Universität München, Germany.

出版信息

Biochem J. 1993 Jul 15;293 ( Pt 2)(Pt 2):501-6. doi: 10.1042/bj2930501.

DOI:10.1042/bj2930501
PMID:8343129
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1134389/
Abstract

We have cloned and sequenced a cDNA encoding a 17.8 kDa subunit of the hydrophobic fragment of complex I from Neurospora crassa. The deduced primary structure of this subunit was partially confirmed by automated Edman degradation of the isolated polypeptide. The sequence data obtained indicate that the 17.8 kDa subunit is made as an extended precursor of 20.8 kDa. Resistance of the polypeptide to alkaline extraction from mitochondrial membranes and the existence of a putative membrane-spanning domain suggests that the 17.8 kDa subunit is an intrinsic (bitopic) membrane protein. The in vitro synthesized precursor of the 17.8 kDa subunit can be efficiently imported into isolated mitochondria, where it is cleaved to the mature species by the metal-dependent matrix-processing peptidase. The in vitro imported mature subunit is found mainly exposed to the mitochondrial intermembrane space. However, a significant fraction of the imported polypeptide acquires the same membrane topology as the endogenous subunit, indicating that correct assembly in the mitochondrial inner membrane did occur.

摘要

我们已经克隆并测序了一个来自粗糙脉孢菌复合体I疏水片段的17.8 kDa亚基的cDNA。通过对分离出的多肽进行自动Edman降解，部分证实了该亚基推导的一级结构。获得的序列数据表明，17.8 kDa亚基最初是以20.8 kDa的延伸前体形式合成的。该多肽对从线粒体膜进行碱性提取具有抗性，并且存在一个假定的跨膜结构域，这表明17.8 kDa亚基是一种内在（双位）膜蛋白。17.8 kDa亚基的体外合成前体可以有效地导入分离的线粒体中，在那里它被金属依赖性的基质加工肽酶切割成成熟形式。体外导入的成熟亚基主要位于线粒体外膜间隙。然而，相当一部分导入的多肽获得了与内源性亚基相同的膜拓扑结构，表明在线粒体内膜中确实发生了正确的组装。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f1e/1134389/078a183330af/biochemj00107-0198-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f1e/1134389/6a91f2f7bbe8/biochemj00107-0195-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f1e/1134389/c1e857368343/biochemj00107-0196-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f1e/1134389/0e60f20a3b42/biochemj00107-0197-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f1e/1134389/9abc327a122b/biochemj00107-0197-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f1e/1134389/078a183330af/biochemj00107-0198-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f1e/1134389/6a91f2f7bbe8/biochemj00107-0195-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f1e/1134389/c1e857368343/biochemj00107-0196-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f1e/1134389/0e60f20a3b42/biochemj00107-0197-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f1e/1134389/9abc327a122b/biochemj00107-0197-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f1e/1134389/078a183330af/biochemj00107-0198-a.jpg

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引用本文的文献

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本文引用的文献

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