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エンドウ トシヤ
ENDO TOSHIYA
遠藤 斗志也 所属 京都産業大学 生命科学部 先端生命科学科 職種 客員教授 |
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| 言語種別 | 英語 |
| 発行・発表の年月 | 2003/04 |
| 形態種別 | 研究論文 |
| 査読 | 査読あり |
| 標題 | Peptide library approach with a disulfide tether to refine the Tom20 recognition motif in mitochondrial presequences |
| 執筆形態 | その他 |
| 掲載誌名 | JOURNAL OF MOLECULAR BIOLOGY |
| 出版社・発行元 | ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD |
| 巻・号・頁 | 328(2),pp.495-504 |
| 著者・共著者 | T Obita,T Muto,T Endo,D Kohda |
| 概要 | Many mitochondrial matrix and inner-membrane proteins are synthesized in the cytosol as precursor proteins with an N-terminal presequence, and are imported into the mitochondria. Although no distinct sequence homology has been found among mitochondrial presequences, Tom20, a general import receptor in the outer mitohcondrial membrane, binds to presequences, and distinguishes mitochondrial proteins from non-mitochonrial proteins. The recently determined structure of the cytosolic domain of Tom20 (DeltaTom20) in a complex with the presequence of rat aldehyde dehydrogenase (ALDH) showed that a short stretch of the presequence forms an amphiphilic helix, and its hydrophobic surface interacts with the hydrophobic-binding groove of Tom20. The following NMR analyses revealed a common five-residue pattern for Tom20 binding in five different presequences. To refine the common amino acid motif for the recognition by Tom20, we introduced a new peptide library approach in this study: we prepared a mixture of ALDH presequence variants, tethered these peptides to DeltaTom20 in a competitive manner by an intermolecular disulfide bond, and determined the relative affinities by MALDI-TOF mass spectrometry. We successfully deduced a refined, common motif for the recognition by Tom20, and found that the segment consisting of residues 14-20 of the ALDH presequence was locally optimized in the sequence space, with respect to Tom20 binding. (C) 2003 Elsevier Science Ltd. All rights reserved. |
| ISSN | 0022-2836 |