| Item type |
文献 / Documents(1) |
| 公開日 |
2018-07-13 |
| アクセス権 |
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アクセス権 |
open access |
| 資源タイプ |
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資源タイプ識別子 |
http://purl.org/coar/resource_type/c_6501 |
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資源タイプ |
journal article |
| 出版社版DOI |
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関連識別子 |
https://doi.org/10.1039/C6OB01014H |
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関連名称 |
10.1039/C6OB01014H |
| 出版タイプ |
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出版タイプ |
AM |
|
出版タイプResource |
http://purl.org/coar/version/c_ab4af688f83e57aa |
| タイトル |
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|
タイトル |
Labelling of Endogenous Target Protein via N–S Acyl Transfer-Mediated Activation of N-Sulfanylethylanilide |
| 著者 |
傳田, 将也
モリサキ, タクヤ
コヒキ, タイキ
ヤマモト, ジュン
サトウ, コウヘイ
佐川, 幾子
猪熊, 翼
佐藤, 陽一
山内, あい子
重永, 章
大髙, 章
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| 抄録 |
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内容記述 |
The ligand-dependent incorporation of a reporter molecule (e.g., fluorescence dye or biotin) onto a endogenous target protein has emerged as an important strategy for elucidating protein function using various affinity-based labelling reagents consisting of reporter, ligand and reactive units. Conventional labelling reagents generally use a weakly activated reactive unit, which can result in the non-specific labelling of proteins in a ligand-independent manner. In this context, the activation of a labelling reagent through a targeted protein-ligand interaction could potentially overcome the problems associated with conventional affinity-based labelling reagents. We hypothesized that this type of protein-ligand-interaction-mediated activation could be accomplished using N-sulfanylethylanilide (SEAlide) as the reactive unit in the labelling reagent. Electrophilically unreactive amide-type SEAlide can be activated by its conversion to the corresponding active thioester in the presence of a phosphate salt, which can act as an acid-base catalyst. It has been suggested that protein surfaces consisting of hydrophilic residues such as amino, carboxyl and imidazole groups could function as acid-base catalysts. We therefore envisioned that a SEAlide-based labelling reagent (SEAL) bearing SEAlide as a reactive unit could be activated through the binding of the SEAL with a target protein. Several SEALs were readily prepared in this study using standard 9-fluorenylmethyloxycarbonyl (Fmoc)-based solid-phase protocols. These SEAL systems were subsequently applied to the ligand-dependent labelling of human carbonic anhydrase (hCA) and cyclooxyganese 1. Although we have not yet obtained any direct evidence for the target protein-mediated activation of the SEAlide unit, our results for the reaction of these SEALs with hCA1 or butylamine indirectly support our hypothesis. The SEALs reported in this study represent valuable new entries to the field of affinity-based labelling reagents and are expected to show great utility in protein labelling. |
| 書誌情報 |
en : Organic & Biomolecular Chemistry
巻 14,
号 26,
p. 6244-6251,
発行日 2016-05-31
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| 収録物ID |
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収録物識別子タイプ |
ISSN |
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収録物識別子 |
14770520 |
| 収録物ID |
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収録物識別子タイプ |
ISSN |
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収録物識別子 |
14770539 |
| 収録物ID |
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収録物識別子タイプ |
NCID |
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収録物識別子 |
AA1168650X |
| 収録物ID |
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収録物識別子タイプ |
NCID |
|
収録物識別子 |
AA12084375 |
| 出版者 |
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|
出版者 |
The Royal Society of Chemistry |
| 権利情報 |
|
|
権利情報 |
© The Royal Society of Chemistry 2016 |
| EID |
|
|
識別子 |
313229 |
| 言語 |
|
|
言語 |
eng |
orbiochem_14_26_6244.pdf (732 KB)
