Aminoacyl-CoAs as probes of condensation domain selectivity in nonribosomal peptide synthesis
Aminoacyl-CoAs as probes of condensation domain selectivity in nonribosomal peptide synthesis
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DOI：
10.1126/science.284.5413.486
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发表时间：
1999-04-16
期刊：
影响因子：
56.9
通讯作者：
Stachelhaus, T
中科院分区：
文献类型：
--
作者：
Belshaw, PJ;Walsh, CT;Stachelhaus, T
在多肽抗生素的非核糖体生物合成中，氨基酸单体被合成酶的腺苷酸化结构域激活，并作为硫酯装载到相邻的载体蛋白结构域上，然后通过合成酶的缩合结构域形成肽键和转移生长链。缩合结构域是否具有编辑功能尚不清楚。氨酰辅酶A（CoA）分子的合成和氨酰-磷酸泛酰巯基乙胺到载体结构域的直接酶促转移允许绕过腺苷酸化结构域编辑功能。该方法用于证明在链起始D-Phe-L-Pro二肽基-酶中间体的形成中，酪氨酸合成酶的第一缩合结构域在供体残基（D-苯丙氨酸）处显示出低选择性，而在受体残基（L-脯氨酸）处显示出较高选择性。
In nonribosomal biosynthesis of peptide antibiotics by multimodular synthetases, amino acid monomers are activated by the adenylation domains of the synthetase and loaded onto the adjacent carrier protein domains as thioesters, then the formation of peptide bonds and translocation of the growing chain are effected by the synthetase's condensation domains. Whether the condensation domains have any editing function has been unknown. Synthesis of aminoacyl-coenzyme A (CoA) molecules and direct enzymatic transfer of aminoacyl-phosphopantetheine to the carrier domains allow the adenylation domain editing function to be bypassed. This method was used to demonstrate that the first condensation domain of tyrocidine synthetase shows low selectivity at the donor residue (D-phenylalanine) and higher selectivity at the acceptor residue (L-proline) in the formation of the chain-initiating D-Phe-L-Pro dipeptidyl-enzyme intermediate.