Involvement of hyp gene products in maturation of the H(2)-sensing [NiFe] hydrogenase of Ralstonia eutropha.
The biosynthesis of [NiFe] hydrogenases is a complex process that requires the function of the Hyp proteins HypA, HypB, HypC, HypD, HypE, HypF, and HypX for assembly of the H(2)-activating [NiFe] site. In this study we examined the maturation of the regulatory hydrogenase (RH) of Ralstonia eutropha. The RH is a H(2)-sensing [NiFe] hydrogenase and is required as a constituent of a signal transduction chain for the expression of two energy-linked [NiFe] hydrogenases. Here we demonstrate that the RH regulatory activity was barely affected by mutations in hypA, hypB, hypC, and hypX and was not substantially diminished in hypD- and hypE-deficient strains. The lack of HypF, however, resulted in a 90% decrease of the RH regulatory activity. Fourier transform infrared spectroscopy and the incorporation of (63)Ni into the RH from overproducing cells revealed that the assembly of the [NiFe] active site is dependent on all Hyp functions, with the exception of HypX. We conclude that the entire Hyp apparatus (HypA, HypB, HypC, HypD, HypE, and HypF) is involved in an efficient incorporation of the [NiFe] center into the RH.
Buhrke
T
T
Institut für Biologie, Humboldt-Universität zu Berlin, 10115 Berlin, Germany.
Bleijlevens
B
B
Albracht
S P
SP
Friedrich
B
B
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D020118:Cupriavidus necator / Q000235:genetics / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D006859:Hydrogen / Q000378:metabolism*; D006864:Hydrogenase / Q000096:biosynthesis* / Q000145:classification; D007501:Iron / Q000378:metabolism; D009154:Mutation; D009532:Nickel / Q000378:metabolism; D010084:Oxidation-Reduction; D010957:Plasmids / Q000235:genetics; D017384:Sequence Deletion; D015398:Signal Transduction
D001426:Bacterial Proteins; D002352:Carrier Proteins; D009532:Nickel; D006859:Hydrogen; D007501:Iron; C043296:nickel-iron hydrogenase; D006864:Hydrogenase; D019204:GTP-Binding Proteins