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
N1 - Funding Information:
We thank S. Löscher and M. Haumann (FU Berlin) for the AAS measurements, I. Zebger (TU Berlin), H. Stosnach and U. Waldschläger (Röntec AG, Berlin) for TXRF analysis, A.L. DeLacey (Madrid, Spain) for the H + /D 2 exchange measurements and J. Stöhr for excellent technical assistance. This work was funded by the Deutsche Forschungsgemeinschaft (SFB498, TP C1) and by the Fonds der Chemischen Industrie. A.K. Jones thanks the Humboldt foundation for a fellowship.
Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
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 - Maturation
KW - Metal center assembly
KW - Ralstonia eutropha
KW - [NiFe] hydrogenase
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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 -