title: Structural basis for the recognition of peptide RJPXD33 by acyltransferases in lipid A biosynthesis.
abstract: UDP-N-acetylglucosamine acyltransferase (LpxA) and UDP-3-O-(acyl)-glucosamine acyltransferase (LpxD) constitute the essential, early acyltransferases of lipid A biosynthesis. Recently, an antimicrobial peptide inhibitor, RJPXD33, was identified with dual affinity for LpxA and LpxD. To gain a fundamental understanding of the molecular basis of inhibitor binding, we determined the crystal structure of LpxA from Escherichia coli in complex with RJPXD33 at 1.9 Å resolutions. Our results suggest that the peptide binds in a unique modality that mimics (R)-β-hydroxyacyl pantetheine binding to LpxA and displays how the peptide binds exclusive of the native substrate, acyl-acyl carrier protein. Acyltransferase binding studies with photo-labile RJPXD33 probes and truncations of RJPXD33 validated the structure and provided fundamental insights for future design of small molecule inhibitors. Overlay of the LpxA-RJPXD33 structure with E. coli LpxD identified a complementary peptide binding pocket within LpxD and serves as a model for further biochemical characterization of RJPXD33 binding to LpxD.
authors.affiliation: From the Department of Medicinal Chemistry, College of Pharmacy, and.
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Jenkins
authors.affiliation: From the Department of Medicinal Chemistry, College of Pharmacy, and.
authors.forename: Kyle A
authors.initials: KA
authors.lastname: Heslip
authors.affiliation: Life Sciences Institute, University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Meagher
authors.affiliation: Life Sciences Institute, University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Jeanne A
authors.initials: JA
authors.lastname: Stuckey
authors.affiliation: From the Department of Medicinal Chemistry, College of Pharmacy, and gdotson@umich.edu.
authors.forename: Garry D
authors.initials: GD
authors.lastname: Dotson
mesh_terms: D000213:Acyl Carrier Protein / Q000378:metabolism; D000217:Acyltransferases / Q000737:chemistry / Q000378:metabolism*; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism*; D054889:Fatty Acid Synthase, Type II / Q000378:metabolism; D008050:Lipid A / Q000096:biosynthesis*; D008958:Models, Molecular; D010204:Pantetheine / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D000213:Acyl Carrier Protein; D029968:Escherichia coli Proteins; D008050:Lipid A; D010455:Peptides; C040157:acpP protein, E coli; D010204:Pantetheine; D000217:Acyltransferases; C050304:acyl-(acyl-carrier-protein)-UDP-N-acetylglucosamine acyltransferase; D054889:Fatty Acid Synthase, Type II
keywords: Acyl Carrier Protein (ACP); Acyltransferase; Antimicrobial Peptide (AMP); Enzyme Inhibitor; Lipid A; Lipopolysaccharide (LPS); LpxA; Peptides; X-ray Crystallography
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