Structure of phosphorylated enzyme I, the phosphoenolpyruvate: sugar phosphotransferase system sugar translocation signal protein

Alexey Teplyakov, Kap Lim, Peng Peng Zhu, Geeta Kapadia, Celia C H Chen, Jennifer M Schwarz, Andrew Howard, Prasad T. Reddy, Alan Peterkofsky, Osnat Herzberg

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

Bacterial transport of many sugars, coupled to their phosphorylation, is carried out by the phosphoenolpyruvate (PEP):sugar phosphotransferase system and involves five phosphoryl group transfer reactions. Sugar translocation initiates with the Mg2+-dependent phosphorylation of enzyme I (EI) by PEP. Crystals of Escherichia coli EI were obtained by mixing the protein with Mg2+ and PEP, followed by oxalate, an EI inhibitor. The crystal structure reveals a dimeric protein where each subunit comprises three domains: a domain that binds the partner PEP:sugar phosphotransferase system protein, HPr; a domain that carries the phosphorylated histidine residue, His-189; and a PEP-binding domain. The PEP-binding site is occupied by Mg2+ and oxalate, and the phosphorylated His-189 is in-line for phosphotransfer to/from the ligand. Thus, the structure represents an enzyme intermediate just after phosphotransfer from PEP and before a conformational transition that brings His-189∼P in proximity to the phosphoryl group acceptor, His-15 of HPr. A model of this conformational transition is proposed whereby swiveling around an α-helical linker disengages the His domain from the PEP-binding domain. Assuming that HPr binds to the HPr-binding domain as observed by NMR spectroscopy of an EI fragment, a rotation around two linker segments orients the His domain relative to the HPr-binding domain so that His-189∼P and His-15 are appropriately stationed for an in-line phosphotransfer reaction.

Original languageEnglish (US)
Pages (from-to)16218-16223
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume103
Issue number44
DOIs
StatePublished - Oct 31 2006
Externally publishedYes

Fingerprint

phosphoenolpyruvate-protein phosphotransferase
Phosphoenolpyruvate
Protein Transport
Phosphoenolpyruvate Sugar Phosphotransferase System
Oxalates
Enzymes
Phosphorylation
Protein Subunits
Enzyme Inhibitors
Histidine
Magnetic Resonance Spectroscopy
Binding Sites

Keywords

  • Phosphorylation
  • Sugar transport
  • X-ray crystallography

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Structure of phosphorylated enzyme I, the phosphoenolpyruvate : sugar phosphotransferase system sugar translocation signal protein. / Teplyakov, Alexey; Lim, Kap; Zhu, Peng Peng; Kapadia, Geeta; Chen, Celia C H; Schwarz, Jennifer M; Howard, Andrew; Reddy, Prasad T.; Peterkofsky, Alan; Herzberg, Osnat.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 103, No. 44, 31.10.2006, p. 16218-16223.

Research output: Contribution to journalArticle

Teplyakov, Alexey ; Lim, Kap ; Zhu, Peng Peng ; Kapadia, Geeta ; Chen, Celia C H ; Schwarz, Jennifer M ; Howard, Andrew ; Reddy, Prasad T. ; Peterkofsky, Alan ; Herzberg, Osnat. / Structure of phosphorylated enzyme I, the phosphoenolpyruvate : sugar phosphotransferase system sugar translocation signal protein. In: Proceedings of the National Academy of Sciences of the United States of America. 2006 ; Vol. 103, No. 44. pp. 16218-16223.
@article{76957da0a6b14eaa923b8fa53ec65b0f,
title = "Structure of phosphorylated enzyme I, the phosphoenolpyruvate: sugar phosphotransferase system sugar translocation signal protein",
abstract = "Bacterial transport of many sugars, coupled to their phosphorylation, is carried out by the phosphoenolpyruvate (PEP):sugar phosphotransferase system and involves five phosphoryl group transfer reactions. Sugar translocation initiates with the Mg2+-dependent phosphorylation of enzyme I (EI) by PEP. Crystals of Escherichia coli EI were obtained by mixing the protein with Mg2+ and PEP, followed by oxalate, an EI inhibitor. The crystal structure reveals a dimeric protein where each subunit comprises three domains: a domain that binds the partner PEP:sugar phosphotransferase system protein, HPr; a domain that carries the phosphorylated histidine residue, His-189; and a PEP-binding domain. The PEP-binding site is occupied by Mg2+ and oxalate, and the phosphorylated His-189 is in-line for phosphotransfer to/from the ligand. Thus, the structure represents an enzyme intermediate just after phosphotransfer from PEP and before a conformational transition that brings His-189∼P in proximity to the phosphoryl group acceptor, His-15 of HPr. A model of this conformational transition is proposed whereby swiveling around an α-helical linker disengages the His domain from the PEP-binding domain. Assuming that HPr binds to the HPr-binding domain as observed by NMR spectroscopy of an EI fragment, a rotation around two linker segments orients the His domain relative to the HPr-binding domain so that His-189∼P and His-15 are appropriately stationed for an in-line phosphotransfer reaction.",
keywords = "Phosphorylation, Sugar transport, X-ray crystallography",
author = "Alexey Teplyakov and Kap Lim and Zhu, {Peng Peng} and Geeta Kapadia and Chen, {Celia C H} and Schwarz, {Jennifer M} and Andrew Howard and Reddy, {Prasad T.} and Alan Peterkofsky and Osnat Herzberg",
year = "2006",
month = "10",
day = "31",
doi = "10.1073/pnas.0607587103",
language = "English (US)",
volume = "103",
pages = "16218--16223",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "44",

}

TY - JOUR

T1 - Structure of phosphorylated enzyme I, the phosphoenolpyruvate

T2 - sugar phosphotransferase system sugar translocation signal protein

AU - Teplyakov, Alexey

AU - Lim, Kap

AU - Zhu, Peng Peng

AU - Kapadia, Geeta

AU - Chen, Celia C H

AU - Schwarz, Jennifer M

AU - Howard, Andrew

AU - Reddy, Prasad T.

AU - Peterkofsky, Alan

AU - Herzberg, Osnat

PY - 2006/10/31

Y1 - 2006/10/31

N2 - Bacterial transport of many sugars, coupled to their phosphorylation, is carried out by the phosphoenolpyruvate (PEP):sugar phosphotransferase system and involves five phosphoryl group transfer reactions. Sugar translocation initiates with the Mg2+-dependent phosphorylation of enzyme I (EI) by PEP. Crystals of Escherichia coli EI were obtained by mixing the protein with Mg2+ and PEP, followed by oxalate, an EI inhibitor. The crystal structure reveals a dimeric protein where each subunit comprises three domains: a domain that binds the partner PEP:sugar phosphotransferase system protein, HPr; a domain that carries the phosphorylated histidine residue, His-189; and a PEP-binding domain. The PEP-binding site is occupied by Mg2+ and oxalate, and the phosphorylated His-189 is in-line for phosphotransfer to/from the ligand. Thus, the structure represents an enzyme intermediate just after phosphotransfer from PEP and before a conformational transition that brings His-189∼P in proximity to the phosphoryl group acceptor, His-15 of HPr. A model of this conformational transition is proposed whereby swiveling around an α-helical linker disengages the His domain from the PEP-binding domain. Assuming that HPr binds to the HPr-binding domain as observed by NMR spectroscopy of an EI fragment, a rotation around two linker segments orients the His domain relative to the HPr-binding domain so that His-189∼P and His-15 are appropriately stationed for an in-line phosphotransfer reaction.

AB - Bacterial transport of many sugars, coupled to their phosphorylation, is carried out by the phosphoenolpyruvate (PEP):sugar phosphotransferase system and involves five phosphoryl group transfer reactions. Sugar translocation initiates with the Mg2+-dependent phosphorylation of enzyme I (EI) by PEP. Crystals of Escherichia coli EI were obtained by mixing the protein with Mg2+ and PEP, followed by oxalate, an EI inhibitor. The crystal structure reveals a dimeric protein where each subunit comprises three domains: a domain that binds the partner PEP:sugar phosphotransferase system protein, HPr; a domain that carries the phosphorylated histidine residue, His-189; and a PEP-binding domain. The PEP-binding site is occupied by Mg2+ and oxalate, and the phosphorylated His-189 is in-line for phosphotransfer to/from the ligand. Thus, the structure represents an enzyme intermediate just after phosphotransfer from PEP and before a conformational transition that brings His-189∼P in proximity to the phosphoryl group acceptor, His-15 of HPr. A model of this conformational transition is proposed whereby swiveling around an α-helical linker disengages the His domain from the PEP-binding domain. Assuming that HPr binds to the HPr-binding domain as observed by NMR spectroscopy of an EI fragment, a rotation around two linker segments orients the His domain relative to the HPr-binding domain so that His-189∼P and His-15 are appropriately stationed for an in-line phosphotransfer reaction.

KW - Phosphorylation

KW - Sugar transport

KW - X-ray crystallography

UR - http://www.scopus.com/inward/record.url?scp=33750802896&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33750802896&partnerID=8YFLogxK

U2 - 10.1073/pnas.0607587103

DO - 10.1073/pnas.0607587103

M3 - Article

C2 - 17053069

AN - SCOPUS:33750802896

VL - 103

SP - 16218

EP - 16223

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 44

ER -