4LXY

Crystal structure WlaRD, a sugar 3N-formyl transferase in the presence of dTDP and 10-N-Formyl-THF

4LXY の概要

• エントリーDOI: 10.2210/pdb4lxy/pdb
• 関連するPDBエントリー: 4LXQ 4LXT 4LXU 4LXX
• 分子名称: WlaRD, a sugar 3N-formyl transferase, N-{4-[{[(6S)-2-amino-4-oxo-3,4,5,6,7,8-hexahydropteridin-6-yl]methyl}(formyl)amino]benzoyl}-L-glutamic acid, THYMIDINE-5'-DIPHOSPHATE, ... (5 entities in total)
• 機能のキーワード: formyltransferase, formylation, transferase
• 由来する生物種: Campylobacter jejuni subsp. jejuni
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 66042.99

構造登録者

Thoden, J.B.,Goneau, M.-F.,Gilbert, M.,Holden, H.M. (登録日: 2013-07-30, 公開日: 2013-08-14, 最終更新日: 2023-09-20)

主引用文献

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

実験手法

X-RAY DIFFRACTION (1.64 Å)