重编程酰基辅酶A泛用转酰基酶的酰基载体蛋白相互作用。
Reprogramming acyl carrier protein interactions of an Acyl-CoA promiscuous trans-acyltransferase.
作者信息
Ye Zhixia, Musiol Ewa M, Weber Tilmann, Williams Gavin J
机构信息
Department of Chemistry, North Carolina State University, Raleigh, NC 27695-8204, USA.
Interfaculty Institute of Microbiology and Infection Medicine, Eberhard Karls University of Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, Germany; German Center for Infection Research (DZIF), Partner Site Tübingen, Tübingen, Germany.
出版信息
Chem Biol. 2014 May 22;21(5):636-46. doi: 10.1016/j.chembiol.2014.02.019. Epub 2014 Apr 10.
Protein interactions between acyl carrier proteins (ACPs) and trans-acting acyltransferase domains (trans-ATs) are critical for regioselective extender unit installation by many polyketide synthases, yet little is known regarding the specificity of these interactions, particularly for trans-ATs with unusual extender unit specificities. Currently, the best-studied trans-AT with nonmalonyl specificity is KirCII from kirromycin biosynthesis. Here, we developed an assay to probe ACP interactions based on leveraging the extender unit promiscuity of KirCII. The assay allows us to identify residues on the ACP surface that contribute to specific recognition by KirCII. This information proved sufficient to modify a noncognate ACP from a different biosynthetic system to be a substrate for KirCII. The findings form a foundation for further understanding the specificity of trans-AT:ACP protein interactions and for engineering modular polyketide synthases to produce analogs.
酰基载体蛋白（ACPs）与反式作用酰基转移酶结构域（trans-ATs）之间的蛋白质相互作用对于许多聚酮合酶进行区域选择性延伸单元的安装至关重要，但对于这些相互作用的特异性，尤其是具有不寻常延伸单元特异性的反式作用酰基转移酶，我们了解甚少。目前，研究最深入的具有非丙二酰特异性的反式作用酰基转移酶是来自奇霉素生物合成的KirCII。在此，我们基于利用KirCII的延伸单元混杂性开发了一种检测方法来探测ACP相互作用。该检测方法使我们能够识别ACP表面上有助于被KirCII特异性识别的残基。这些信息被证明足以将来自不同生物合成系统的非同源ACP修饰为KirCII的底物。这些发现为进一步理解反式作用酰基转移酶与ACP的蛋白质相互作用的特异性以及工程化模块化聚酮合酶以生产类似物奠定了基础。
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