2N5T

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

Summary for 2N5T

• Entry DOI: 10.2210/pdb2n5t/pdb
• NMR Information: BMRB: 25731
• Descriptor: Phosphoenolpyruvate-protein phosphotransferase (1 entity in total)
• Functional Keywords: transferase
• Biological source: Escherichia coli
• Total number of polymer chains: 2
• Total formula weight: 127271.29

Authors

Venditti, V.,Schwieters, C.D.,Grishaev, A.,Clore, G. (deposition date: 2015-07-28, release date: 2015-09-02, Last modification date: 2024-05-01)

Primary citation

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

Experimental method

SOLUTION NMR
SOLUTION SCATTERING