A chemical reporter for protein AMPylation.

A chemical reporter for protein AMPylation.
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DOI:
10.1021/ja205137d
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发表时间:
2011-11-02
影响因子:
15
通讯作者:
Hang, Howard C.
Hang, Howard C.
中科院分区:
化学1区
文献类型:
--
作者:
Grammel, Markus;Phi Luong;Orth, Kim;Hang, Howard C.

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蛋白ampyation是一种新兴的翻译后修饰,在细菌发病和细胞生物学中起着关键作用。具有ampyylation活性的酶,被称为ampyators,已经在几种细菌病原体和真核生物中被发现。为了促进这种独特修饰的研究,我们开发了一种炔基化学报告基因,用于检测和鉴定蛋白质AMPylation底物。ampyylation底物与炔基报告物的共价功能化代替5 ' -单磷酸腺苷基(AMP),允许它们随后与叠氮化物荧光染料或亲和富集标签进行生物正交连接。我们表明,该化学报告基因通过一系列ampylator转移到其同源蛋白底物上,并允许快速检测和鉴定AMPylated底物。
Protein AMPylation is an emerging posttranslational modification, which plays key roles in bacterial pathogenesis and cell biology. Enzymes with AMPylation activity, referred to as AMPylators, have been identified in several bacterial pathogens and eukaryotes. To facilitate the study of this unique modification, we developed an alkynyl chemical reporter for detection and identification of protein AMPylation substrates. Covalent functionalization of AMPylation substrates with the alkynyl reporter in lieu of adenylyl 5′-monophosphate (AMP) allows their subsequent bioorthogonal ligation with azide-fluorescent dyes or affinity enrichment tags. We show that this chemical reporter is transferred by a range of AMPylators onto their cognate protein substrates and allows rapid detection and identification of AMPylated substrates.
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