The H2-sensing complex of Ralstonia eutropha: interaction between a regulatory [NiFe] hydrogenase and a histidine protein kinase.
Two [NiFe] hydrogenases enable the proteobacterium Ralstonia eutropha H16 to grow on molecular hydrogen as the sole energy source. A third [NiFe] hydrogenase (RH) acts as an H2 sensor in a multiple component signal transduction chain that controls hydrogenase gene transcription. The RH forms a dimeric heterodimer (HoxBC)2 in which HoxC contains the H2-sensing active site and HoxB the electron-transferring components including an organic, not yet identified redox cofactor. This oligomer forms a tight complex with the histidine protein kinase HoxJ. Both the sensor and the kinase were analysed by mutagenesis for functional domains that are instrumental in H2 signal transmission. A mutant deleted for a C-terminal peptide of 55 amino acids in HoxB lost its H2-sensing ability but still catalysed H2 oxidation. The mutant protein failed to form the dimeric heterodimer and a complex with HoxJ. The organic redox cofactor was no longer detectable in the truncated sensor. H2 sensing was also abolished by deletion of the PAS domain of HoxJ, indicating that this domain is involved in signal transduction. A truncated version of HoxJ consisting of only the input domain of the kinase was still capable of forming a complex with the RH. Mass determination of the purified HoxJ protein revealed that the kinase forms a homotetramer. The unique oligomeric structure of the H2-sensing complex with respect to its regulatory function is discussed.
Institut für Biologie/Mikrobiologie, Humboldt-Universität zu Berlin, Chausseestr 117, 10115 Berlin, Germany.
Thorsten
T
Buhrke
Oliver
O
Lenz
Antje
A
Porthun
Bärbel
B
Friedrich
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020118:Cupriavidus necator / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D019281:Dimerization; D015964:Gene Expression Regulation, Bacterial; D000071677:Histidine Kinase; D006859:Hydrogen / Q000378:metabolism*; D006864:Hydrogenase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009532:Nickel / Q000378:metabolism; D010957:Plasmids; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D009532:Nickel; D006859:Hydrogen; C043296:nickel-iron hydrogenase; D006864:Hydrogenase; D011494:Protein Kinases; D000071677:Histidine Kinase; C104634:HoxJ protein, bacteria