Title: Structural and Biochemical Analysis of Protein-Protein Interactions Between the Acyl-Carrier Protein and Product Template Domain.
Authors: Barajas, Jesus F; Finzel, Kara; Valentic, Timothy R; Shakya, Gaurav; Gamarra, Nathan; Martinez, Delsy; Meier, Jordan L; Vagstad, Anna L; Newman, Adam G; Townsend, Craig A; Burkart, Michael D; Tsai, Shiou-Chuan
Published In Angew Chem Int Ed Engl, (2016 10 10)
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.
PubMed ID: 27653519
MeSH Terms: Acyl Carrier Protein/chemistry*; Acyl Carrier Protein/metabolism; Molecular Conformation; Molecular Docking Simulation; Polyketides/chemistry*; Polyketides/metabolism; Protein Binding; Protein Conformation