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Journal of Bacteriology, July 2002, p. 3879-3885, Vol. 184, No. 14
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.14.3879-3885.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Network of Hydrogenase Maturation in Escherichia coli: Role of Accessory Proteins HypA and HybF

Michaela Hube, Melanie Blokesch, and August Böck*

Department Biologie I, Mikrobiologie, Universität München, D-80638 Munich, Germany

Received 4 March 2002/ Accepted 26 April 2002

We have studied the roles of the auxiliary protein HypA and of its homolog HybF in hydrogenase maturation. A mutation in hypA leads to the nearly complete blockade of maturation solely of hydrogenase 3 whereas a lesion in hybF drastically but not totally reduces maturation and activity of isoenzymes 1 and 2. The residual level of matured enzymes in the hybF mutant was shown to be due to the function of HypA; HybF, conversely, was responsible for a minimal residual activity of hydrogenase 3 in the mutant hypA strain. Accordingly, a hypA {Delta}hybF double mutant was completely blocked in the maturation process. However, the inclusion of high nickel concentrations in the medium could restore limited activity of all three hydrogenases. The results of this study and of previous work (M. Blokesch, A. Magalon, and A. Böck, J. Bacteriol. 189:2817-2822, 2001) show that the maturation of the three functional hydrogenases from Escherichia coli is intimately connected via the activity of proteins HypA and HypC and of their homologs HybF and HybG, respectively. The results also support the suggestion of Olson et al. (J. W. Olson, N. S. Mehta, and R. J. Maier, Mol. Microbiol. 39:176-182, 2001) that HypA cooperates with HypB in the insertion of nickel into the precursor of the large hydrogenase subunit. Whereas HypA is predominantly involved in the maturation of hydrogenase 3, HybF takes over its function in the maturation of isoenzymes 1 and 2.

* Corresponding author. Mailing address: Department Biologie I, Mikrobiologie, University of Munich, Maria-Ward-Straße 1a, D-80638 Munich, Germany. Phone: 49(0)89-2180-6120. Fax: 49(0)89-2180-6122. E-mail: august.boeck{at}lrz.uni-muenchen.de.

Journal of Bacteriology, July 2002, p. 3879-3885, Vol. 184, No. 14
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.14.3879-3885.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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