6T6V

Glu-494-Ala inactive monomer of a quinol dependent Nitric Oxide Reductase (qNOR) from Alcaligenes xylosoxidans

6T6V の概要

• エントリーDOI: 10.2210/pdb6t6v/pdb
• EMDBエントリー: 10387
• 分子名称: Nitric oxide reductase subunit B, PROTOPORPHYRIN IX CONTAINING FE, CALCIUM ION (3 entities in total)
• 機能のキーワード: monomer, proton transfer, nitric oxide, oxidoreductase
• 由来する生物種: Alcaligenes xylosoxydans xylosoxydans
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 86039.01

構造登録者

Gopalasingam, C.C.,Johnson, R.M.,Antonyuk, S.V.,Muench, S.P.,Hasnain, S.S. (登録日: 2019-10-19, 公開日: 2020-04-01, 最終更新日: 2024-05-22)

主引用文献

Jamali, M.A.M.,Gopalasingam, C.C.,Johnson, R.M.,Tosha, T.,Muramoto, K.,Muench, S.P.,Antonyuk, S.V.,Shiro, Y.,Hasnain, S.S.
The active form of quinol-dependent nitric oxide reductase fromNeisseria meningitidisis a dimer.
Iucrj, 7:404-415, 2020

PubMed Abstract: is carried by nearly a billion humans, causing developmental impairment and over 100 000 deaths a year. A quinol-dependent nitric oxide reductase (qNOR) plays a critical role in the survival of the bacterium in the human host. X-ray crystallographic analyses of qNOR, including that from (qNOR) reported here at 3.15 Å resolution, show monomeric assemblies, despite the more active dimeric sample being used for crystallization. Cryo-electron microscopic analysis of the same chromatographic fraction of qNOR, however, revealed a dimeric assembly at 3.06 Å resolution. It is shown that zinc (which is used in crystallization) binding near the dimer-stabilizing TMII region contributes to the disruption of the dimer. A similar destabilization is observed in the monomeric (∼85 kDa) cryo-EM structure of a mutant (Glu494Ala) qNOR from the opportunistic pathogen () , which primarily migrates as a monomer. The monomer-dimer transition of qNORs seen in the cryo-EM and crystallographic structures has wider implications for structural studies of multimeric membrane proteins. X-ray crystallographic and cryo-EM structural analyses have been performed on the same chromatographic fraction of qNOR to high resolution. This represents one of the first examples in which the two approaches have been used to reveal a monomeric assembly and a dimeric assembly in vitrified cryo-EM grids. A number of factors have been identified that may trigger the destabilization of helices that are necessary to preserve the integrity of the dimer. These include zinc binding near the entry of the putative proton-transfer channel and the preservation of the conformational integrity of the active site. The mutation near the active site results in disruption of the active site, causing an additional destabilization of helices (TMIX and TMX) that flank the proton-transfer channel helices, creating an inert monomeric enzyme.

PubMed: 32431824
DOI: 10.1107/S2052252520003656

実験手法

ELECTRON MICROSCOPY (4.5 Å)