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ツゲ ヒデアキ
TSUGE HIDEAKI
津下 英明 所属 京都産業大学 生命科学部 先端生命科学科 職種 教授 |
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| 言語種別 | 英語 |
| 発行・発表の年月 | 2015/03 |
| 形態種別 | 研究論文 |
| 査読 | 査読あり |
| 標題 | Structural Basis for Action of the External Chaperone for a Propeptide-deficient Serine Protease from Aeromonas sobria. |
| 執筆形態 | その他 |
| 掲載誌名 | The Journal of Biological Chemistry |
| 出版社・発行元 | AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC |
| 巻・号・頁 | 290(17),pp.11130-11143 |
| 担当区分 | 最終著者,責任著者 |
| 著者・共著者 | Kobayashi H,Yoshida T,Miyakawa T,Tashiro M,Okamoto K,Yamanaka H,Tanokura M,Tsuge H |
| 概要 | Subtilisin-like proteases are broadly expressed in organisms ranging from bacteria to mammals. During maturation of these enzymes, N-terminal propeptides function as intramolecular chaperones, assisting the folding of their catalytic domains. However, we have identified an exceptional case, the serine protease from Aeromonas sobria (ASP), that lacks a propeptide. Instead, ORF2, a protein encoded just downstream of asp, appears essential for proper ASP folding. The mechanism by which ORF2 functions remains an open question, because it shares no sequence homology with any known intramolecular propeptide or other protein. Here we report the crystal structure of the ORF2-ASP complex and the solution structure of free ORF2. ORF2 consists of three regions: an N-terminal extension, a central body, and a C-terminal tail. Together, the structure of the central body and the C-terminal tail is similar to that of the intramolecular propeptide. The N-terminal extension, which is not seen in other subtilisin-like enzymes, is intrinsically disordered but forms some degree of secondary structure upon binding ASP. Wealso show that C-terminal (Delta C1 and Delta C5) or N-terminal (Delta N43 and Delta N64) deletion eliminates the ability of ORF2 to function as a chaperone. Characterization of the maturation of ASP with ORF2 showed that folding occurs in the periplasmic space and is followed by translocation into extracellular space and dissociation from ORF2, generating active ASP. Finally, a PSI-BLAST search revealed that operons encoding subtilases and their external chaperones are widely distributed among Gram-negative bacteria, suggesting that ASP and its homologs form a novel family of subtilases having an external chaperone. |
| DOI | 10.1074/jbc.M114.622852 |
| ISSN | 0021-9258/1083-351X |
| PMID | 25784551 |
| Put Code(ORCID) | 15839291 |
| PermalinkURL | http://europepmc.org/abstract/med/25784551 |