Analysis of recognition elements for mitochondrial processing peptidase using artificial amino acids: roles of the intervening portion and proximal arginine.
Analysis of recognition elements for mitochondrial processing peptidase using artificial amino acids: roles of the intervening portion and proximal arginine.
复制标题
使用人工氨基酸分析线粒体加工肽酶的识别元件：中间部分和近端精氨酸的作用。
DOI：
10.1093/oxfordjournals.jbchem.a022529
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
A. Ito
中科院分区：
文献类型：
--
作者：
K. Moriwaki;T. Ogishima;A. Ito
最近，我们利用模拟苹果酸脱氢酶延伸肽的合成肽，证明了相对于裂解位点近端和远端的氨基酸残基是线粒体加工肽酶识别底物的关键决定因素[Niidome等人]。(1994)J.Biol.化学。269、24719-24722)。虽然近端的精氨酸无一例外地位于-2位置，但远端残基的位置在线粒体前体蛋白中是不同的。在近端和远端残基之间，脯氨酸和/或甘氨酸存在于大多数线粒体前体蛋白中，它们被认为在肽酶对底物的特异性识别中发挥作用。为了阐明介入部分的作用，我们在远端和近端残基之间引入了非天然氨基酸[2-(2-氨基乙氧基)乙酸]。我们还分析了近端精氨酸的功能成分，用不同的精氨酸或赖氨酸类似物取代残基。动力学研究的结果表明，中间部分应该是灵活的，以便于有效的加工，并且近端精氨酸的胍基是通过氢键和离子键被肽酶识别的。
We recently demonstrated, using synthetic peptides modeled on the extension peptide of malate dehydrogenase, that amino acid residues present at the proximal and distal positions relative to the cleavage site are critical determinants for the recognition of substrates by mitochondrial processing peptidase [Niidome et al. (1994) J. Biol. Chem. 269, 24719-24722). While the proximal arginine is unexceptionally located at the -2 position, the position of the distal residue varies among mitochondrial precursor proteins. Between the proximal and distal residues, proline and/or glycine are present in most mitochondrial precursor proteins, and they are considered to play a role in the specific recognition of a substrate by the peptidase. To elucidate the role of the intervening portion, we introduced a non-natural amino acid [2-(2-aminoethoxy)acetic acid] between the distal and proximal residues. We also analyzed the functional elements in the proximal arginine by replacing the residue with various arginine or lysine analogs. The results of kinetic studies indicated that the intervening portion should be flexible for efficient processing, and that the guanidino group of the proximal arginine is recognized by the peptidase through hydrogen and ionic bonds.