title: Reprogramming acyl carrier protein interactions of an Acyl-CoA promiscuous trans-acyltransferase.
abstract: 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.
authors.affiliation: Department of Chemistry, North Carolina State University, Raleigh, NC 27695-8204, USA.
authors.forename: Zhixia
authors.initials: Z
authors.lastname: Ye
authors.affiliation: 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.
authors.forename: Ewa M
authors.initials: EM
authors.lastname: Musiol
authors.affiliation: 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.
authors.forename: Tilmann
authors.initials: T
authors.lastname: Weber
authors.affiliation: Department of Chemistry, North Carolina State University, Raleigh, NC 27695-8204, USA. Electronic address: gavin_williams@ncsu.edu.
authors.forename: Gavin J
authors.initials: GJ
authors.lastname: Williams
mesh_terms: D000213:Acyl Carrier Protein / Q000378:metabolism*; D000214:Acyl Coenzyme A / Q000378:metabolism*; D000217:Acyltransferases / Q000378:metabolism*; D015394:Molecular Structure; D011728:Pyridones / Q000378:metabolism
chemical_list: D000213:Acyl Carrier Protein; D000214:Acyl Coenzyme A; D011728:Pyridones; D000217:Acyltransferases; C008536:mocimycin
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