Arginine residues in the extension peptide are required for cleavage of a precursor by mitochondrial processing peptidase. Demonstration using synthetic peptide as a substrate.
Arginine residues in the extension peptide are required for cleavage of a precursor by mitochondrial processing peptidase. Demonstration using synthetic peptide as a substrate.
复制标题
线粒体加工肽酶裂解前体需要延伸肽中的精氨酸残基。
DOI：
--
复制
发表时间：
1994
影响因子：
4.8
通讯作者：
Akio Ito
中科院分区：
文献类型：
--
作者：
T. Niidome;S. Kitada;K. Shimokata;T. Ogishima;Akio Ito
线粒体加工肽酶（MPP）特异性识别多种线粒体前体蛋白并正确切割延伸肽。为了确定MPP特异性识别所需的所有延伸肽的共同结构，我们基于前苹果酸脱氢酶的延伸肽的氨基酸序列合成了不同链长和氨基酸序列的各种寡肽，并确定了切割反应的动力学参数。有效切割的肽的最小长度为16个氨基酸残基，分别由从切割位点到氨基和羧基末端侧的11个和5个残基组成。肽中的两组碱性氨基酸，相对于切割位点的位置-10的远端精氨酸残基和位置-3和-2的近端精氨酸残基是有效水解所必需的。在这两个中，位置-2的残基对于有效切割比位置-3的残基更重要，并且不能被赖氨酸残基取代。用赖氨酸替换远端精氨酸对切割没有影响。我们的研究表明，使用具有适当长度的肽是必不可少的MPP切割反应进行动力学分析，并在位置-2的精氨酸残基的切割位点是必要的识别和切割的延伸肽。
Mitochondrial processing peptidase (MPP) specifically recognizes a large variety of mitochondrial precursor proteins and correctly cleaves off the extension peptides. To determine the structure common to all the extension peptides that is required for specific recognition by MPP, we synthesized various oligopeptides of different chain lengths and amino acid sequences, based on the amino acid sequence of the extension peptide of pre-malate dehydrogenase, and determined kinetic parameters of the cleavage reactions. The minimal length of peptides for effective cleavage was 16 amino acid residues consisting of 11 and 5 residues from the cleavage site to the amino- and carboxyl-terminal sides, respectively. Two sets of basic amino acids in the peptide, the distal arginine residue at position -10 and the proximal ones at positions -3 and -2 relative to the cleavage site, were necessary for effective hydrolysis. Of these two, the residue at position -2 was more important for effective cleavage than the one at position -3 and could not be replaced by a lysine residue. The replacement of the distal arginine by lysine had no effect on the cleavage. Our study demonstrates that use of peptides with the proper length is essential for performing kinetic analyses on the cleavage reaction by MPP and that an arginine residue at position -2 to the cleavage site is necessary for the recognition and cleavage of the extension peptide.