Structure and malonyl CoA-ACP transacylase binding of streptomyces coelicolor fatty acid synthase acyl carrier protein.
Malonylation of an acyl carrier protein (ACP) by malonyl Coenzyme A-ACP transacylase (MCAT) is fundamental to bacterial fatty acid biosynthesis. Here, we report the structure of the Steptomyces coelicolor (Sc) fatty acid synthase (FAS) ACP and studies of its binding to MCAT. The carrier protein adopts an alpha-helical bundle structure common to other known carrier proteins. The Sc FAS ACP shows close structural homology with other fatty acid ACPs and less similarity with Sc actinorhodin (act) polyketide synthase (PKS) ACP where the orientation of helix I differs. NMR experiments were used to map the binding of ACP to MCAT. This data suggests that Sc FAS ACP interacts with MCAT through the negatively charged helix II of ACP, consistent with proposed models for ACP recognition by other FAS enzymes. Differential roles for residues at the interface are demonstrated using site-directed mutagenesis and in vitro assays. MCAT has been suggested, moreover, to participate in bacterial polyketide synthesis in vivo. We demonstrate that the affinity of the polyketide synthase ACP for MCAT is lower than that of the FAS ACP. Mutagenesis of homologous helix II residues on the polyketide synthase ACP suggests that the PKS ACP may bind to MCAT in a different manner than the FAS counterpart.
Arthur
Christopher J
CJ
School of Chemistry, University of Bristol, Cantock's Close, Bristol BS8 1TS, UK.
Williams
Christopher
C
Pottage
Katherine
K
Płoskoń
Eliza
E
Findlow
Stuart C
SC
Burston
Steven G
SG
Simpson
Thomas J
TJ
Crump
Matthew P
MP
Crosby
John
J
D051080:Acyl-Carrier Protein S-Malonyltransferase / Q000737:chemistry* / Q000378:metabolism*; D064429:Fatty Acid Synthases / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D048372:Streptomyces coelicolor / Q000737:chemistry* / Q000378:metabolism*
D051080:Acyl-Carrier Protein S-Malonyltransferase; D064429:Fatty Acid Synthases