Structural and Biochemical Analysis of Protein–Protein Interactions Between the Acyl‐Carrier Protein and Product Template Domain

• DOI: 10.1002/anie.201605401
• Publication Date: 2016-09-22T08:19:03Z
• Authors: Jesus F. Barajas; Kara Finzel; Timothy R. Valentic; Gaurav Shakya; Nathan Gamarra; Delsy Martinez; Jordan L. Meier; Anna L. Vagstad; Adam G. Newman; Craig A. Townsend; Michael D. Burkart; Shiou‐Chuan Tsai
• Keywords: 0301 basic medicine; 570; 0303 health sciences; Protein Conformation; polyketide synthase; Organic Chemistry; Molecular Conformation; Bioengineering; acyl-carrier protein; product template domain; Molecular Docking Simulation; 03 medical and health sciences; Chemical sciences; Polyketides; Chemical Sciences; Acyl Carrier Protein; crosslinking; Infection; Protein Binding

AbstractIn 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.