Structure of a sugar N -formyltransferase from campylobacter jejuni

  1. Get@NRC: Structure of a sugar N -formyltransferase from campylobacter jejuni (Opens in a new window)
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Journal titleBiochemistry
Pages61146126; # of pages: 13
SubjectAmino acid residues; Campylobacter jejuni; Carbonyl carbon; Catalytic efficiencies; Enzymatic activities; Gram-negative bacteria; Tetrahydrofolates; X-ray structure; Amino acids; Biochemistry; Carbon; Catalysis; Enzymes; Bacteria; amino acid; carbon; carbonyl derivative; folinic acid; galactose; glucose; histidine; n formyltransferase; O antigen; sugar; tetrahydrofolic acid; thymidine diphosphate; transferase; unclassified drug; article; biosynthesis; Campylobacter jejuni; catalysis; enzyme activity; enzyme assay; enzyme structure; mutation; nonhuman; outer membrane; priority journal; X ray
AbstractThe O-antigens, which are components of the outer membranes of Gram-negative bacteria, are responsible for the wide species variations seen in nature and are thought to play a role in bacterial virulence. They often contain unusual dideoxysugars such as 3,6-dideoxy-3-formamido-d-glucose (Qui3NFo). Here, we describe a structural and functional investigation of the protein C8J-1081 from Campylobacter jejuni 81116, which is involved in the biosynthesis of Qui3NFo. Specifically, the enzyme, hereafter referred to as WlaRD, catalyzes the N-formylation of dTDP-3,6-dideoxy-3-amino-d-glucose (dTDP-Qui3N) using N10-formyltetrahydrofolate as the carbon source. For this investigation, seven X-ray structures of WlaRD, in complexes with various dTDP-linked sugars and cofactors, were determined to resolutions of 1.9 Å or better. One of the models, with bound N10-formyltetrahydrofolate and dTDP, represents the first glimpse of an N-formyltransferase with its natural cofactor. Another model contains the reaction products, tetrahydrofolate and dTDP-Qui3NFo. In combination, the structures provide snapshots of the WlaRD active site before and after catalysis. On the basis of these structures, three amino acid residues were targeted for study: Asn 94, His 96, and Asp 132. Mutations of any of these residues resulted in a complete loss of enzymatic activity. Given the position of His 96 in the active site, it can be postulated that it functions as the active site base to remove a proton from the sugar amino group as it attacks the carbonyl carbon of the N-10 formyl group of the cofactor. Enzyme assays demonstrate that WlaRD is also capable of utilizing dTDP-3,6-dideoxy-3-amino-d-galactose (dTDP-Fuc3N) as a substrate, albeit at a much reduced catalytic efficiency. © 2013 American Chemical Society.
Publication date
AffiliationNational Research Council Canada (NRC-CNRC); Human Health Therapeutics (HHT-TSH)
Peer reviewedYes
NPARC number21269870
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Record identifierd5786577-2561-4ca9-919d-573c7d994d4a
Record created2013-12-13
Record modified2016-05-09
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