Specific enrichment of nonribosomal peptide synthetase module by an affinity probe for adenylation domains
Specific enrichment of nonribosomal peptide synthetase module by an affinity probe for adenylation domains
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DOI：
10.1016/j.bmcl.2013.12.082
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发表时间：
2014-02-01
影响因子：
2.7
通讯作者：
Kakeya, Hideaki
中科院分区：
文献类型：
--
作者：
Ishikawa, Fumihiro;Kakeya, Hideaki
我们针对非核糖体肽合成酶(NRPS)-聚酮合成酶（PKS）杂种和NRPSs中腺苷化(A)结构域的亲和探针的开发，可以促进含有A结构域模块的富集、鉴定和定量。一种5'- o -氨基磺酰基腺苷（AMS）的不可水解类似物单磷酸腺苷（AMP）已被报道为设计具有紧密结合腺苷化酶抑制剂的支架。在这里，我们描述了A域亲和探针的应用。我们的合成探针是一种生物素化的L-Phe- ams (L-Phe- ams -biotin)，专门针对NRPS模块中的a结构域，在重组NRPS酶系统和整个蛋白质组中激活L-Phe为氨基酰腺苷酸中间体。(C) 2014 Elsevier Ltd.版权所有。
We targeted the development of an affinity probe for adenylation (A) domains that can facilitate enrichment, identification, and quantification of A domain-containing modules in nonribosomal peptide synthetase (NRPS)-polyketide synthase (PKS) hybrids and NRPSs. A 5'-O-sulfamoyladenosine (AMS) non-hydrolyzable analogue of adenosine monophosphate (AMP) has been reported as a scaffold for the design of inhibitors exhibiting tight binding of adenylation enzymes. Here we describe the application of an affinity probe for A domains. Our synthetic probe, a biotinylated L-Phe-AMS (L-Phe-AMS-biotin) specifically targets the A domains in NRPS modules that activates L-Phe to an aminoacyladenylate intermediate in both recombinant NRPS enzyme systems and whole proteomes. (C) 2014 Elsevier Ltd. All rights reserved.