通过用于腺苷化结构域的亲和探针实现非核糖体肽合成酶模块的特异性富集。
Specific enrichment of nonribosomal peptide synthetase module by an affinity probe for adenylation domains.
作者信息
Ishikawa Fumihiro, Kakeya Hideaki
机构信息
Department of System Chemotherapy and Molecular Sciences, Division of Bioinformatics and Chemical Genomics, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
出版信息
Bioorg Med Chem Lett. 2014 Feb 1;24(3):865-9. doi: 10.1016/j.bmcl.2013.12.082. Epub 2013 Dec 25.
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.
我们致力于开发一种用于腺苷化（A）结构域的亲和探针，该探针能够促进非核糖体肽合成酶（NRPS）-聚酮合酶（PKS）杂合体和NRPS中含A结构域模块的富集、鉴定和定量。5'-O-氨磺酰腺苷（AMS），一种单磷酸腺苷（AMP）的不可水解类似物，已被报道作为设计与腺苷化酶紧密结合的抑制剂的支架。在此，我们描述了一种用于A结构域的亲和探针的应用。我们的合成探针，生物素化的L-苯丙氨酸-AMS（L-Phe-AMS-生物素），在重组NRPS酶系统和完整蛋白质组中，特异性靶向NRPS模块中的A结构域，该结构域将L-苯丙氨酸激活为氨酰腺苷酸中间体。
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