The Structure of Hydrogenase-2 from Escherichia coli: Implications for H2 -Driven Proton Pumping

Stephen E.  Beaton; Rhiannon M. Evans; Alexander J. Finney; Claire M. Lamont; Fraser A. Armstrong; Frank Sargent; Stephen B. Carr

doi:10.1042/BCJ20180053

The Structure of Hydrogenase-2 from Escherichia coli: Implications for H₂ -Driven Proton Pumping

Stephen E. Beaton
, Rhiannon M. Evans
, Alexander J. Finney
, Claire M. Lamont
, Fraser A. Armstrong (Lead / Corresponding author)
, Frank Sargent (Lead / Corresponding author)
, Stephen B. Carr (Lead / Corresponding author)

Molecular Microbiology

Research output: Contribution to journal › Article › peer-review

54 Citations (Scopus)

355 Downloads (Pure)

Abstract

Under anaerobic conditions Escherichia coli is able to metabolize molecular hydrogen via the action of several [NiFe]-hydrogenase enzymes. Hydrogenase-2, which is typically present in cells at low levels during anaerobic respiration, is a periplasmic-facing membrane-bound complex that functions as a proton pump to convert energy from H2 oxidation into a proton gradient; consequently, its structure is of great interest. Empirically, the complex consists of a tightlybound core catalytic module, comprising large (HybC) and small (HybO) subunits, which is attached to an Fe-S protein (HybA) and an integral membrane protein, HybB. To date, efforts to gain a more detailed picture have been thwarted by low native expression levels of hydrogenase-2 and the labile interaction between HybOC and HybA/HybB subunits. In this paper we describe a new over-expression system that has facilitated determination of highresolution crystal structures of HybOC and, hence, a prediction of the quaternary structure of the HybOCAB complex.

Original language	English
Pages (from-to)	1353-1370
Number of pages	18
Journal	Biochemical Journal
Volume	475
Issue number	7
Early online date	19 Mar 2018
DOIs	https://doi.org/10.1042/BCJ20180053
Publication status	Published - 16 Apr 2018

Keywords

Hydrogenase
Metalloenzyme
Escherichia coli
Iron-sulphur protein
Protein structure

ASJC Scopus subject areas

Molecular Biology
Biochemistry
Cell Biology

Access to Document

10.1042/BCJ20180053

Final Published VersionFinal published version, 8.26 MBLicence: CC BY

Cite this

@article{f26821bd983d45c4ab507aea8f30f97b,

title = "The Structure of Hydrogenase-2 from Escherichia coli: Implications for H2 -Driven Proton Pumping",

abstract = "Under anaerobic conditions Escherichia coli is able to metabolize molecular hydrogen via the action of several [NiFe]-hydrogenase enzymes. Hydrogenase-2, which is typically present in cells at low levels during anaerobic respiration, is a periplasmic-facing membrane-bound complex that functions as a proton pump to convert energy from H2 oxidation into a proton gradient; consequently, its structure is of great interest. Empirically, the complex consists of a tightlybound core catalytic module, comprising large (HybC) and small (HybO) subunits, which is attached to an Fe-S protein (HybA) and an integral membrane protein, HybB. To date, efforts to gain a more detailed picture have been thwarted by low native expression levels of hydrogenase-2 and the labile interaction between HybOC and HybA/HybB subunits. In this paper we describe a new over-expression system that has facilitated determination of highresolution crystal structures of HybOC and, hence, a prediction of the quaternary structure of the HybOCAB complex.",

keywords = "Hydrogenase, Metalloenzyme, Escherichia coli, Iron-sulphur protein, Protein structure ",

author = "Beaton, \{Stephen E.\} and Evans, \{Rhiannon M.\} and Finney, \{Alexander J.\} and Lamont, \{Claire M.\} and Armstrong, \{Fraser A.\} and Frank Sargent and Carr, \{Stephen B.\}",

note = "This research was funded in Oxford by the Biotechnology and Biological Sciences Research Council (BBSRC) Grant BB/N006321/1, and in Dundee by BBSRC EASTBIO Doctoral Training Partnership awards 1510231 and 1280664. FAA is a Royal Society Wolfson Research Merit Award holder. SEB is grateful to the Dakota Foundation for the award of a Holaday Scholarship.",

year = "2018",

month = apr,

day = "16",

doi = "10.1042/BCJ20180053",

language = "English",

volume = "475",

pages = "1353--1370",

journal = "Biochemical Journal",

issn = "0264-6021",

publisher = "Portland Press",

number = "7",

}

TY - JOUR

T1 - The Structure of Hydrogenase-2 from Escherichia coli

T2 - Implications for H2 -Driven Proton Pumping

AU - Beaton, Stephen E.

AU - Evans, Rhiannon M.

AU - Finney, Alexander J.

AU - Lamont, Claire M.

AU - Armstrong, Fraser A.

AU - Sargent, Frank

AU - Carr, Stephen B.

N1 - This research was funded in Oxford by the Biotechnology and Biological Sciences Research Council (BBSRC) Grant BB/N006321/1, and in Dundee by BBSRC EASTBIO Doctoral Training Partnership awards 1510231 and 1280664. FAA is a Royal Society Wolfson Research Merit Award holder. SEB is grateful to the Dakota Foundation for the award of a Holaday Scholarship.

PY - 2018/4/16

Y1 - 2018/4/16

N2 - Under anaerobic conditions Escherichia coli is able to metabolize molecular hydrogen via the action of several [NiFe]-hydrogenase enzymes. Hydrogenase-2, which is typically present in cells at low levels during anaerobic respiration, is a periplasmic-facing membrane-bound complex that functions as a proton pump to convert energy from H2 oxidation into a proton gradient; consequently, its structure is of great interest. Empirically, the complex consists of a tightlybound core catalytic module, comprising large (HybC) and small (HybO) subunits, which is attached to an Fe-S protein (HybA) and an integral membrane protein, HybB. To date, efforts to gain a more detailed picture have been thwarted by low native expression levels of hydrogenase-2 and the labile interaction between HybOC and HybA/HybB subunits. In this paper we describe a new over-expression system that has facilitated determination of highresolution crystal structures of HybOC and, hence, a prediction of the quaternary structure of the HybOCAB complex.

AB - Under anaerobic conditions Escherichia coli is able to metabolize molecular hydrogen via the action of several [NiFe]-hydrogenase enzymes. Hydrogenase-2, which is typically present in cells at low levels during anaerobic respiration, is a periplasmic-facing membrane-bound complex that functions as a proton pump to convert energy from H2 oxidation into a proton gradient; consequently, its structure is of great interest. Empirically, the complex consists of a tightlybound core catalytic module, comprising large (HybC) and small (HybO) subunits, which is attached to an Fe-S protein (HybA) and an integral membrane protein, HybB. To date, efforts to gain a more detailed picture have been thwarted by low native expression levels of hydrogenase-2 and the labile interaction between HybOC and HybA/HybB subunits. In this paper we describe a new over-expression system that has facilitated determination of highresolution crystal structures of HybOC and, hence, a prediction of the quaternary structure of the HybOCAB complex.

KW - Hydrogenase

KW - Metalloenzyme

KW - Escherichia coli

KW - Iron-sulphur protein

KW - Protein structure

U2 - 10.1042/BCJ20180053

DO - 10.1042/BCJ20180053

M3 - Article

C2 - 29555844

SN - 0264-6021

VL - 475

SP - 1353

EP - 1370

JO - Biochemical Journal

JF - Biochemical Journal

IS - 7

ER -