Chemoenzymatic exchange of phosphopantetheine on protein and peptide
Chemoenzymatic exchange of phosphopantetheine on protein and peptide
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DOI：
10.1039/c3sc53154f
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发表时间：
2014-03-01
期刊：
影响因子：
8.4
通讯作者：
Burkart, Michael D.
中科院分区：
文献类型：
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作者：
Kosa, Nicolas M.;Pham, Kevin M.;Burkart, Michael D.
对新的酰基载体蛋白水解酶（AcpH, EC 3.1.4.14）同源物的评价表明，从载体蛋白水解磷酸蚁氨酸的底物选择性和动力学参数存在显著差异。对来自初级和次级代谢途径的载体蛋白的评估显示，II型脂肪酸合成酶的酰基载体蛋白（ACP）底物总体上更受青睐，而聚酮合成酶ACP和来自非核糖体肽合成酶的肽基载体蛋白（PCP）的活性也各不相同。我们还展示了AcpP和11聚肽底物YbbR的这些同源物的动力学参数。这些发现使得这三类天然产物合酶载体蛋白以及完整和最小融合蛋白结构的完全可逆标记成为可能。
Evaluation of new acyl carrier protein hydrolase (AcpH, EC 3.1.4.14) homologs from proteobacteria and cyanobacteria reveals significant variation in substrate selectivity and kinetic parameters for phosphopantetheine hydrolysis from carrier proteins. Evaluation with carrier proteins from both primary and secondary metabolic pathways reveals an overall preference for acyl carrier protein (ACP) substrates from type II fatty acid synthases, as well as variable activity for polyketide synthase ACPs and peptidyl carrier proteins (PCP) from non-ribosomal peptide synthases. We also demonstrate the kinetic parameters of these homologs for AcpP and the 11-mer peptide substrate YbbR. These findings enable the fully reversible labeling of all three classes of natural product synthase carrier proteins as well as full and minimal fusion protein constructs.