A model system for [NiFe] hydrogenase maturation studies: Purification of an active site-containing hydrogenase large subunit without small subunit

Gordon Winter, Thorsten Buhrke, Oliver Lenz, Anne Katherine Jones, Michael Forgber, Bärbel Friedrich

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

The large subunit HoxC of the H2-sensing [NiFe] hydrogenase from Ralstonia eutropha was purified without its small subunit. Two forms of HoxC were identified. Both forms contained iron but only substoichiometric amounts of nickel. One form was a homodimer of HoxC whereas the second also contained the Ni-Fe site maturation proteins HypC and HypB. Despite the presence of the Ni-Fe active site in some of the proteins, both forms, which lack the Fe-S clusters normally present in hydrogenases, cannot activate hydrogen. The incomplete insertion of nickel into the Ni-Fe site provides direct evidence that Fe precedes Ni in the course of metal center assembly.

Original languageEnglish
Pages (from-to)4292-4296
Number of pages5
JournalFEBS Letters
Volume579
Issue number20
DOIs
Publication statusPublished - Aug 15 2005

Fingerprint

Hydrogenase
Nickel
Purification
Catalytic Domain
Cupriavidus necator
Hydrogen
Proteins
Iron
Metals
nickel-iron hydrogenase

Keywords

  • [NiFe] hydrogenase
  • Maturation
  • Metal center assembly
  • Ralstonia eutropha

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

A model system for [NiFe] hydrogenase maturation studies : Purification of an active site-containing hydrogenase large subunit without small subunit. / Winter, Gordon; Buhrke, Thorsten; Lenz, Oliver; Jones, Anne Katherine; Forgber, Michael; Friedrich, Bärbel.

In: FEBS Letters, Vol. 579, No. 20, 15.08.2005, p. 4292-4296.

Research output: Contribution to journalArticle

Winter, Gordon ; Buhrke, Thorsten ; Lenz, Oliver ; Jones, Anne Katherine ; Forgber, Michael ; Friedrich, Bärbel. / A model system for [NiFe] hydrogenase maturation studies : Purification of an active site-containing hydrogenase large subunit without small subunit. In: FEBS Letters. 2005 ; Vol. 579, No. 20. pp. 4292-4296.
@article{a18e4e48358f43d29f55ba307b690a97,
title = "A model system for [NiFe] hydrogenase maturation studies: Purification of an active site-containing hydrogenase large subunit without small subunit",
abstract = "The large subunit HoxC of the H2-sensing [NiFe] hydrogenase from Ralstonia eutropha was purified without its small subunit. Two forms of HoxC were identified. Both forms contained iron but only substoichiometric amounts of nickel. One form was a homodimer of HoxC whereas the second also contained the Ni-Fe site maturation proteins HypC and HypB. Despite the presence of the Ni-Fe active site in some of the proteins, both forms, which lack the Fe-S clusters normally present in hydrogenases, cannot activate hydrogen. The incomplete insertion of nickel into the Ni-Fe site provides direct evidence that Fe precedes Ni in the course of metal center assembly.",
keywords = "[NiFe] hydrogenase, Maturation, Metal center assembly, Ralstonia eutropha",
author = "Gordon Winter and Thorsten Buhrke and Oliver Lenz and Jones, {Anne Katherine} and Michael Forgber and B{\"a}rbel Friedrich",
year = "2005",
month = "8",
day = "15",
doi = "10.1016/j.febslet.2005.06.064",
language = "English",
volume = "579",
pages = "4292--4296",
journal = "FEBS Letters",
issn = "0014-5793",
publisher = "Elsevier",
number = "20",

}

TY - JOUR

T1 - A model system for [NiFe] hydrogenase maturation studies

T2 - Purification of an active site-containing hydrogenase large subunit without small subunit

AU - Winter, Gordon

AU - Buhrke, Thorsten

AU - Lenz, Oliver

AU - Jones, Anne Katherine

AU - Forgber, Michael

AU - Friedrich, Bärbel

PY - 2005/8/15

Y1 - 2005/8/15

N2 - The large subunit HoxC of the H2-sensing [NiFe] hydrogenase from Ralstonia eutropha was purified without its small subunit. Two forms of HoxC were identified. Both forms contained iron but only substoichiometric amounts of nickel. One form was a homodimer of HoxC whereas the second also contained the Ni-Fe site maturation proteins HypC and HypB. Despite the presence of the Ni-Fe active site in some of the proteins, both forms, which lack the Fe-S clusters normally present in hydrogenases, cannot activate hydrogen. The incomplete insertion of nickel into the Ni-Fe site provides direct evidence that Fe precedes Ni in the course of metal center assembly.

AB - The large subunit HoxC of the H2-sensing [NiFe] hydrogenase from Ralstonia eutropha was purified without its small subunit. Two forms of HoxC were identified. Both forms contained iron but only substoichiometric amounts of nickel. One form was a homodimer of HoxC whereas the second also contained the Ni-Fe site maturation proteins HypC and HypB. Despite the presence of the Ni-Fe active site in some of the proteins, both forms, which lack the Fe-S clusters normally present in hydrogenases, cannot activate hydrogen. The incomplete insertion of nickel into the Ni-Fe site provides direct evidence that Fe precedes Ni in the course of metal center assembly.

KW - [NiFe] hydrogenase

KW - Maturation

KW - Metal center assembly

KW - Ralstonia eutropha

UR - http://www.scopus.com/inward/record.url?scp=23644455994&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=23644455994&partnerID=8YFLogxK

U2 - 10.1016/j.febslet.2005.06.064

DO - 10.1016/j.febslet.2005.06.064

M3 - Article

C2 - 16061234

AN - SCOPUS:23644455994

VL - 579

SP - 4292

EP - 4296

JO - FEBS Letters

JF - FEBS Letters

SN - 0014-5793

IS - 20

ER -