2N5T

Ensemble solution structure of the phosphoenolpyruvate-Enzyme I complex from the bacterial phosphotransferase system

2N5T の概要

• エントリーDOI: 10.2210/pdb2n5t/pdb
• NMR情報: BMRB: 25731
• 分子名称: Phosphoenolpyruvate-protein phosphotransferase (1 entity in total)
• 機能のキーワード: transferase
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 127271.29

構造登録者

Venditti, V.,Schwieters, C.D.,Grishaev, A.,Clore, G. (登録日: 2015-07-28, 公開日: 2015-09-02, 最終更新日: 2024-05-01)

主引用文献

Venditti, V.,Schwieters, C.D.,Grishaev, A.,Clore, G.M.
Dynamic equilibrium between closed and partially closed states of the bacterial Enzyme I unveiled by solution NMR and X-ray scattering.
Proc.Natl.Acad.Sci.USA, 112:11565-11570, 2015

PubMed Abstract: Enzyme I (EI) is the first component in the bacterial phosphotransferase system, a signal transduction pathway in which phosphoryl transfer through a series of bimolecular protein-protein interactions is coupled to sugar transport across the membrane. EI is a multidomain, 128-kDa homodimer that has been shown to exist in two conformational states related to one another by two large (50-90°) rigid body domain reorientations. The open conformation of apo EI allows phosphoryl transfer from His189 located in the N-terminal domain α/β (EIN(α/β)) subdomain to the downstream protein partner bound to the EIN(α) subdomain. The closed conformation, observed in a trapped phosphoryl transfer intermediate, brings the EIN(α/β) subdomain into close proximity to the C-terminal dimerization domain (EIC), thereby permitting in-line phosphoryl transfer from phosphoenolpyruvate (PEP) bound to EIC to His189. Here, we investigate the solution conformation of a complex of an active site mutant of EI (H189A) with PEP. Simulated annealing refinement driven simultaneously by solution small angle X-ray scattering and NMR residual dipolar coupling data demonstrates unambiguously that the EI(H189A)-PEP complex exists in a dynamic equilibrium between two approximately equally populated conformational states, one corresponding to the closed structure and the other to a partially closed species. The latter likely represents an intermediate in the open-to-closed transition.

PubMed: 26305976
DOI: 10.1073/pnas.1515366112

実験手法

SOLUTION NMR
SOLUTION SCATTERING