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Solution structure and conformational dynamics of a doublet acyl carrier protein from prodigiosin biosynthesis

DOI: 10.1021/acs.biochem.0c00830 DOI Help

Authors: Thitapa Thongkawphueak (Kasetsart University) , Ashley J. Winter (University of Bristol) , Christopher Williams (University of Bristol) , Hannah J. Maple (University of Bristol) , Siriwat Soontaranon (Synchrotron Light Research Institute) , Chonthicha Kaewhan (Synchrotron Light Research Institute) , Dominic J. Campopiano (University of Edinburgh) , Matthew P. Crump (University of Bristol) , Pakorn Wattana-amorn (Kasetsart University)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Biochemistry

State: Published (Approved)
Published: January 2021
Diamond Proposal Number(s): 23269

Abstract: The acyl carrier protein (ACP) is an indispensable component of both fatty acid and polyketide synthases and is primarily responsible for delivering acyl intermediates to enzymatic partners. At present, increasing numbers of multidomain ACPs have been discovered with roles in molecular recognition of trans-acting enzymatic partners as well as increasing metabolic flux. Further structural information is required to provide insight into their function, yet to date, the only high-resolution structure of this class to be determined is that of the doublet ACP (two continuous ACP domains) from mupirocin synthase. Here we report the solution nuclear magnetic resonance (NMR) structure of the doublet ACP domains from PigH (PigH ACP1-ACP2), which is an enzyme that catalyzes the formation of the bipyrrolic intermediate of prodigiosin, a potent anticancer compound with a variety of biological activities. The PigH ACP1-ACP2 structure shows each ACP domain consists of three conserved helices connected by a linker that is partially restricted by interactions with the ACP1 domain. Analysis of the holo (4′-phosphopantetheine, 4′-PP) form of PigH ACP1-ACP2 by NMR revealed conformational exchange found predominantly in the ACP2 domain reflecting the inherent plasticity of this ACP. Furthermore, ensemble models obtained from SAXS data reveal two distinct conformers, bent and extended, of both apo (unmodified) and holo PigH ACP1-ACP2 mediated by the central linker. The bent conformer appears to be a result of linker–ACP1 interactions detected by NMR and might be important for intradomain communication during the biosynthesis. These results provide new insights into the behavior of the interdomain linker of multiple ACP domains that may modulate protein–protein interactions. This is likely to become an increasingly important consideration for metabolic engineering in prodigiosin and other related biosynthetic pathways.

Journal Keywords: Peptides and proteins; Monomers; X-ray scattering; Chemical structure; Conformation

Subject Areas: Biology and Bio-materials, Chemistry


Instruments: B21-High Throughput SAXS

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