title: Structural and Biochemical Analysis of Protein-Protein Interactions Between the Acyl-Carrier Protein and Product Template Domain.
abstract: In fungal non-reducing polyketide synthases (NR-PKS) the acyl-carrier protein (ACP) carries the growing polyketide intermediate through iterative rounds of elongation, cyclization and product release. This process occurs through a controlled, yet enigmatic coordination of the ACP with its partner enzymes. The transient nature of ACP interactions with these catalytic domains imposes a major obstacle for investigation of the influence of protein-protein interactions on polyketide product outcome. To further our understanding about how the ACP interacts with the product template (PT) domain that catalyzes polyketide cyclization, we developed the first mechanism-based crosslinkers for NR-PKSs. Through in vitro assays, in silico docking and bioinformatics, ACP residues involved in ACP-PT recognition were identified. We used this information to improve ACP compatibility with non-cognate PT domains, which resulted in the first gain-of-function ACP with improved interactions with its partner enzymes. This advance will aid in future combinatorial biosynthesis of new polyketides.
authors.affiliation: Department of Molecular Biology and Biochemistry, Chemistry, and Pharmaceutical Sciences, University of California, Irvine, Irvine, CA, 92697, USA.
authors.forename: Jesus F
authors.initials: JF
authors.lastname: Barajas
authors.affiliation: Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, 92093, USA.
authors.forename: Kara
authors.initials: K
authors.lastname: Finzel
authors.affiliation: Department of Molecular Biology and Biochemistry, Chemistry, and Pharmaceutical Sciences, University of California, Irvine, Irvine, CA, 92697, USA.
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Valentic
authors.affiliation: Department of Molecular Biology and Biochemistry, Chemistry, and Pharmaceutical Sciences, University of California, Irvine, Irvine, CA, 92697, USA.
authors.forename: Gaurav
authors.initials: G
authors.lastname: Shakya
authors.affiliation: Department of Molecular Biology and Biochemistry, Chemistry, and Pharmaceutical Sciences, University of California, Irvine, Irvine, CA, 92697, USA.
authors.forename: Nathan
authors.initials: N
authors.lastname: Gamarra
authors.affiliation: Department of Molecular Biology and Biochemistry, Chemistry, and Pharmaceutical Sciences, University of California, Irvine, Irvine, CA, 92697, USA.
authors.forename: Delsy
authors.initials: D
authors.lastname: Martinez
authors.affiliation: Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, 92093, USA.
authors.forename: Jordan L
authors.initials: JL
authors.lastname: Meier
authors.affiliation: Department of Chemistry, The Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, USA.
authors.forename: Anna L
authors.initials: AL
authors.lastname: Vagstad
authors.affiliation: Department of Chemistry, The Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, USA.
authors.forename: Adam G
authors.initials: AG
authors.lastname: Newman
authors.affiliation: Department of Chemistry, The Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, USA.
authors.forename: Craig A
authors.initials: CA
authors.lastname: Townsend
authors.affiliation: Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, 92093, USA. mburkart@ucsd.edu.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Burkart
authors.affiliation: Department of Molecular Biology and Biochemistry, Chemistry, and Pharmaceutical Sciences, University of California, Irvine, Irvine, CA, 92697, USA. sctsai@uci.edu.
authors.forename: Shiou-Chuan
authors.initials: SC
authors.lastname: Tsai
mesh_terms: D000213:Acyl Carrier Protein / Q000737:chemistry* / Q000378:metabolism; D008968:Molecular Conformation; D062105:Molecular Docking Simulation; D061065:Polyketides / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation
chemical_list: D000213:Acyl Carrier Protein; D061065:Polyketides
keywords: acyl-carrier protein; crosslinking; polyketide synthase; product template domain