title: A photocrosslinking assay for reporting protein interactions in polyketide and fatty acid synthases.
abstract: Understanding protein-protein interactions that occur between ACP and KS domains of polyketide synthases and fatty acid synthases is critical to improving the scope and efficiency of combinatorial biosynthesis efforts aimed at producing non-natural polyketides. Here, we report a facile strategy for rapidly reporting such ACP-KS interactions based on the incorporation of an amino acid with photocrosslinking functionality. Crucially, this photocrosslinking strategy can be applied to any polyketide or fatty acid synthase regardless of substrate specificity, and can be adapted to a high-throughput format for directed evolution studies.
authors.affiliation: Department of Chemistry, North Carolina State University, Raleigh, NC 27695 USA.
authors.forename: Zhixia
authors.initials: Z
authors.lastname: Ye
authors.forename: Morgan
authors.initials: M
authors.lastname: Bair
authors.forename: Hemant
authors.initials: H
authors.lastname: Desai
authors.forename: Gavin J
authors.initials: GJ
authors.lastname: Williams
mesh_terms: D000213:Acyl Carrier Protein / Q000737:chemistry / Q000378:metabolism; D018388:Codon, Terminator; D003432:Cross-Linking Reagents / Q000737:chemistry* / Q000378:metabolism; D064429:Fatty Acid Synthases / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D048630:Polyketide Synthases / Q000737:chemistry* / Q000378:metabolism; D061065:Polyketides / Q000378:metabolism; D011487:Protein Conformation
chemical_list: D000213:Acyl Carrier Protein; D018388:Codon, Terminator; D003432:Cross-Linking Reagents; D061065:Polyketides; D048630:Polyketide Synthases; D064429:Fatty Acid Synthases