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UDP-GalNAc : Polypeptide N-acetylgalactosaminyltransferase 1(GalNAc-T1)の構造活性相関 : Gal/GalNAc-Tモチーフ中の保存された芳香族アミノ酸残基の機能解析
http://hdl.handle.net/10965/463
http://hdl.handle.net/10965/463dd1211fd-5ccf-4342-833d-4837dccf96d9
| 名前 / ファイル | ライセンス | アクション |
|---|---|---|
AHSUSK_NSS_33_45.pdf (254.1 kB)
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| Item type | 紀要論文 / Departmental Bulletin Paper(1) | |||||
|---|---|---|---|---|---|---|
| 公開日 | 2017-09-30 | |||||
| タイトル | ||||||
| タイトル | UDP-GalNAc : Polypeptide N-acetylgalactosaminyltransferase 1(GalNAc-T1)の構造活性相関 : Gal/GalNAc-Tモチーフ中の保存された芳香族アミノ酸残基の機能解析 | |||||
| 言語 | ja | |||||
| タイトル | ||||||
| タイトル | Structure-function Relationship of UDP-GalNAc : Polypeptide N-Acetylgalactosaminyltransferase 1 (GalNAc-T1) : Function of Conserved Aromatic Residues in the Gal/GalNAc-T Motif of GalNAc-T1 | |||||
| 言語 | en | |||||
| 言語 | ||||||
| 言語 | eng | |||||
| キーワード | ||||||
| 主題Scheme | Other | |||||
| 主題 | ムチン型O-グリコシレーション | |||||
| キーワード | ||||||
| 主題Scheme | Other | |||||
| 主題 | GalNAc 転移酵素 | |||||
| キーワード | ||||||
| 主題Scheme | Other | |||||
| 主題 | Gal/GalNAc-T モチーフ | |||||
| キーワード | ||||||
| 主題Scheme | Other | |||||
| 主題 | 構造活性相関 | |||||
| キーワード | ||||||
| 主題Scheme | Other | |||||
| 主題 | 部位特異的変異 | |||||
| キーワード | ||||||
| 言語 | en | |||||
| 主題Scheme | Other | |||||
| 主題 | mucin-type O-glycosylation | |||||
| キーワード | ||||||
| 言語 | en | |||||
| 主題Scheme | Other | |||||
| 主題 | GalNAc-transferase | |||||
| キーワード | ||||||
| 言語 | en | |||||
| 主題Scheme | Other | |||||
| 主題 | Gal/GalNAc-T motif | |||||
| キーワード | ||||||
| 言語 | en | |||||
| 主題Scheme | Other | |||||
| 主題 | structure-function relationship | |||||
| キーワード | ||||||
| 言語 | en | |||||
| 主題Scheme | Other | |||||
| 主題 | site-directed mutagenesis | |||||
| 資源タイプ | ||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
| 資源タイプ | departmental bulletin paper | |||||
| 著者 |
天野, 麻理
× 天野, 麻理 |
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| 抄録 | ||||||
| 内容記述タイプ | Abstract | |||||
| 内容記述 | UDP-GalNAc: polypeptide N-acetylgalactosaminyltransferases (GalNAc-transferases) catalyze an initial step of mucin-type carbohydrate biosynthesis by transferring GalNAc to a hydroxylamino acid on acceptor proteins. It is reported that the GalNAc-transferases catalyze two distinguishable reactions; one is the initial reaction that transfers GalNAc to unglycosylated acceptors, and the other is the follow-up reaction that glycosylates partially glycosylated acceptors. Here, I report the structure-function relationship of the Gal/GalNAc-T motif, which occurs in both the GalNAc-transferases and the β4-galactosyltransferases. I investigated the role of the six conserved aromatic residues in this motif in GalNAc-T1, one of the most ubiquitous isozymes, and evaluated the role of each aromatic residue, using site-directed mutagenesis, as follows. 1) Tyr302 and Phe325 might not be important for reaction. 2) Phe303 and Tyr309 might be modestly involved in the interactions with both the sugar-donor and sugaracceptor substrates. 3) Trp328 and Trp316 are important sites for enzymatic activity, since the mutations at either site resulted in complete inactivation of the enzyme. I then investigated the role of Trp328 and Trp316 in more detail by generating additional mutants and analyzing the resultant mutants kinetically. Their role was deduced as follows. 1) Trp328 is an essential residue for the activity, since all the mutants generated were inactive and Trp328 is conserved in all the isozymes cloned to date. 2) An aromatic residue is required for the activity at the position of 316. 3) Trp316 is an important binding site with acceptors, primarily involved in the initial glycosylation of acceptor polypeptides. This study first pinpointed the location of the amino acid residue associated with the initial activity of the GalNAc-transferases. | |||||
| 内容記述 | ||||||
| 内容記述タイプ | Other | |||||
| 内容記述 | 1. Introduction 2. Materials and Methods 2.1 Site-directed mutagenesis of soluble rat recombinant GalNAc-T1 2.2 Expression of P-ΔN42 and mutant P-ΔN42 in COS7 cells 2.3 GalNAc-transferase assay 3. Results 3.1 Mutagenesis of the conserved aromatic residues in the Gal/GalNAc-T motif 3.2 Kinetic analysis of the mutant enzymes 3.3 Mutagenesis of Trp316 and Trp328 in the Gal/GalNAc-T motif 3.4 Kinetic analysis of Trp316 mutant enzymes 3.5 Trp316 is involved in the initial glycosylation of the acceptor polypeptides 3.6 Involvement of Trp316 in the binding of acceptor peptides with a single acceptor site 4. Discussion |
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| 書誌情報 |
京都産業大学論集. 自然科学系列 巻 33, p. 45-59, 発行日 2004-03 |
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| 出版者 | ||||||
| 出版者 | 京都産業大学 | |||||
| ISSN | ||||||
| 収録物識別子タイプ | PISSN | |||||
| 収録物識別子 | 1348-3323 | |||||
| 書誌レコードID | ||||||
| 収録物識別子タイプ | NCID | |||||
| 収録物識別子 | AA11923897 | |||||
| 著者版フラグ | ||||||
| 出版タイプ | VoR | |||||
| 出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||
