7RML

Neisseria meningitidis Methylenetetrahydrofolate reductase in complex with FAD

7RML の概要

• エントリーDOI: 10.2210/pdb7rml/pdb
• 分子名称: 5,10-methylenetetrahydrofolate reductase, FLAVIN-ADENINE DINUCLEOTIDE (3 entities in total)
• 機能のキーワード: flavoprotein, oxidoreductase
• 由来する生物種: Neisseria meningitidis serogroup B (strain MC58)
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 104142.56

構造登録者

Pederick, J.L.,Wegener, K.L.,Salaemae, W.,Bruning, J.B. (登録日: 2021-07-27, 公開日: 2022-11-02, 最終更新日: 2023-10-25)

主引用文献

Pantong, W.,Pederick, J.L.,Maenpuen, S.,Tinikul, R.,Jayapalan, J.J.,Jovcevski, B.,Wegener, K.L.,Bruning, J.B.,Salaemae, W.
Biochemical and structural characterization of meningococcal methylenetetrahydrofolate reductase.
Protein Sci., 32:e4654-e4654, 2023

PubMed Abstract: Methylenetetrahydrofolate reductase (MTHFR) is a key metabolic enzyme in colonization and virulence of Neisseria meningitidis, a causative agent of meningococcal diseases. Here, the biochemical and structural properties of MTHFR from a virulent strain of N. meningitidis serogroup B (NmMTHFR) were characterized. Unlike other orthologs, NmMTHFR functions as a unique homohexamer, composed of three homo-dimerization partners, as shown in our 2.7 Å resolution crystal structure. Six active sites were formed solely within monomers and located away from the oligomerization interfaces. Flavin adenine dinucleotide cofactor formed hydrogen bonds with conserved sidechains, positioning its isoalloxazine ring adjacent to the overlapping binding sites of nicotinamide adenine dinucleotide (NADH) coenzyme and CH -H folate substrate. NmMTHFR utilized NADH (K  = 44 μM) as an electron donor in the NAD(P)H-CH -H folate oxidoreductase assay, but not nicotinamide adenine dinucleotide phosphate (NADPH) which is the donor required in human MTHFR. In silico analysis and mutagenesis studies highlighted the significant difference in orientation of helix α7A (Phe215-Thr225) with that in the human enzyme. The extended sidechain of Met221 on helix α7A plays a role in stabilizing the folded structure of NADH in the hydrophobic box. This supports the NADH specificity by restricting the phosphate group of NADPH that causes steric clashes with Glu26. The movement of Met221 sidechain allows the CH -H folate substrate to bind. The unique topology of its NADH and CH -H folate binding pockets makes NmMTHFR a promising drug target for the development of new antimicrobial agents that may possess reduced off-target side effects.

PubMed: 37165541
DOI: 10.1002/pro.4654

実験手法

X-RAY DIFFRACTION (2.7 Å)