Campus Units
Chemistry
Document Type
Article
Publication Version
Published Version
Publication Date
8-4-2015
Journal or Book Title
Proceedings of the National Academy of Sciences of the United States of America
Volume
112
Issue
37
First Page
11565
Last Page
11570
DOI
10.1073/pnas.1515366112
Abstract
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 (EINα/β) subdomain to the downstream protein partner bound to the EINα subdomain. The closed conformation, observed in a trapped phosphoryl transfer intermediate, brings the EINα/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
Rights
Works produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted.
Language
en
File Format
application/pdf
Recommended Citation
Venditti, Vincenzo; Schwieters, Charles D.; Grishaev, Alexander; and Clore, G. Marius, "Dynamic equilibrium between closed and partially closed states of the bacterial Enzyme I unveiled by solution NMR and X-ray scattering" (2015). Chemistry Publications. 1000.
https://lib.dr.iastate.edu/chem_pubs/1000
Comments
This is an article from Venditti, Vincenzo, Charles D. Schwieters, Alexander Grishaev, and G. Marius Clore. "Dynamic equilibrium between closed and partially closed states of the bacterial Enzyme I unveiled by solution NMR and X-ray scattering." Proceedings of the National Academy of Sciences 112, no. 37 (2015): 11565-11570. doi: 10.1073/pnas.1515366112. Posted with permission.