4LXU

dTdp-Fuc3N and 5-N-Formyl-THF

Summary for 4LXU

• Entry DOI: 10.2210/pdb4lxu/pdb
• Related: 4LXQ 4LXT 4LXX 4LXY 4LY0 4LY3
• Descriptor: WlaRD, a sugar 3N-formyl transferase, N-{[4-({[(6R)-2-amino-5-formyl-4-oxo-1,4,5,6,7,8-hexahydropteridin-6-yl]methyl}amino)phenyl]carbonyl}-L-glutamic acid, 3[N-MORPHOLINO]PROPANE SULFONIC ACID, ... (8 entities in total)
• Functional Keywords: fmt, formyltransferase, n-10-formyl-thf, formylation, transferase
• Biological source: Campylobacter jejuni subsp. jejuni
• Total number of polymer chains: 2
• Total formula weight: 67120.96

Authors

Thoden, J.B.,Goneau, M.-F.,Gilbert, M.,Holden, H.M. (deposition date: 2013-07-30, release date: 2013-08-14, Last modification date: 2023-09-20)

Primary citation

Thoden, J.B.,Goneau, M.F.,Gilbert, M.,Holden, H.M.
Structure of a sugar N-formyltransferase from Campylobacter jejuni.
Biochemistry, 52:6114-6126, 2013

PubMed Abstract: The 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 N(10)-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 N(10)-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.

PubMed: 23898784
DOI: 10.1021/bi4009006

Experimental method

X-RAY DIFFRACTION (1.4 Å)