Characterization of hydrogenase

Ma K, R Weiss, MWW Adams (2000) Characterization of hydrogenase II from the hyperthermophilic archaeon Pyrococcus furiosus and assessment of its role in sulfur reduction. J Bacteriol 182 1864-1871. 

An outline of the biochemical methods used for isolation and biochemical characterization of hydrogenases is given here. Further details are given in a review (Cammack et al. 1994). 

Bryant, F. O. and Adams, M. W. (1989) Characterization of hydrogenase from the hyperther-mophilic archaebacterium, Pyrococcus furiosus. J. Biol. Chem., 264, 5070-9. 

Spectroscopic Characterization of Hydrogenase Enzymes Electron paramagnetic resonance... 

Tamagnini, P., Oxelfelt, F., Salema, R. and Lindblad, P. (1995). Immunological characterization of hydrogenases in the nitrogen-fixing cyanobacterium Nostoc sp. strain PCC 73102. Curr. Microbiol. 31, 102-107. 

Ueno, Y., Kurano, N., Miyachi, S. (1999). Purification and characterization of hydrogenase from the marine green alga, Chlorococcum littorale. FEBS Letters, 443, 144-148. 

Especially for this latter class of hydrogenases, great effort has been devoted to the purification and the characterization of the metal centers involved, using biochemical, genetic, spectroscopic (IR, EPR, Mossbauer, MCD, EXAFS, and mass spectrometry), and crystallographic techniques 152, 165, 166). 

R. L. Csaki. Bodrossy, T. Hanczar, J. C. Murrell, K. L. Kovacs (2001) Molecular characterization of a membrane bound hydrogenase in the methanotroph Methylococcus capsulatus (Bath). FEMSMicrobiol. Lett., 205 203-207... 

Isolation and characterization of the [Fe]-hydrogenase gene from C. moewusii... 

T. Happe, JD. Naber (1993) Isolation, characterization and N-terminal amino acid sequence of hydrogenase from the green alga Chlamydomonas reinhardtii. Eur. J. Biochem., 214 475-481... 

M. Winkler, B. Heil, T. Happe (2002) Isolation and molecular characterization of the [Fe]-hydrogenase from the unicellular green alga Chlorella fiisca. Biochem. Biophys. Acta, 1576 330-334... 

As described above, the combination of EPR and Mossbauer spectroscopies, when applied to carefully prepared parallel samples, enables a detailed characterization of all the redox states of the clusters present in the enzyme. Once the characteristic spectroscopic properties of each cluster are identified, the determination of their midpoint redox potentials is an easy task. Plots of relative amounts of each species (or some characteristic intensive property) as a function of the potential can be fitted to Nernst equations. In the case of the D. gigas hydrogenase it was determined that those midpoint redox potentials (at pFi 7.0) were —70 mV for the [3Fe-4S] [3Fe-4S]° and —290 and —340mV for each of the [4Fe-4S]> [4Fe-4S] transitions. 

Berber, Y., Fauque, G. D., LeGall, J., Choi, E. S., Peck, H. D. Jr and Lespinat, P. A. (1987) Inhibition studies of three classes of Desulfovibrio hydrogenase Application to the further characterization of the multiple hydrogenases found in Desulfovibrio vulgaris Hildenborough. Biochem. Biophys. Res. Commun., 146, 147-53. 

Black, L. K., Fu, C. and Maier, R. J. (1994) Sequences and characterization of hupU and hupV genes of Bradyrhizobium japonicum encoding a possible nickel-sensing complex involved in hydrogenase expression./. BacterioL, 176, 7102-6. 

Fan, C. L., Teixeira, M., Moura, J., Moura, I., Huynh, B. H., Legall, J., Peck, H. D. and Hoffman, B. M. (1991) Detection and characterization of exchangeable protons bound to the hydrogen-activation nickel site of Desulfovibrio gigas hydrogenase - a H-1 and H-2 Q-band ENDOR study./. Amer. Chem. Soc., 113, 1-24... 

Lutz, S., Jacobi, A., Schlensog, V., Bohm, R., Sawers, G. and Bock, A. (1991) Molecular characterization of an operon (hyp) necessary for the activity of the three hydrogenase isoenzymes in Escherichia coli. Mol. Microbiol., 5, 123-35. 

Michel, R., Massanz, C., Kostka, S., Richter, M. and Fiebig, K. (1995) Biochemical characterization of the 8-hydroxy-5-deazaflavin-reactive hydrogenase from Methanosarcina barkeri Fusaro. Eur. J. Biochem., 233, 727-35. 

Muth, E., Morschel, E. and Klein, A. (1987) Purification and characterization of an 8-hydroxy-5-deazaflavin- reducing hydrogenase from the archaebacterium Methanococcus voltae. Eur. J. Biochem., 169, 571-7. 

Osterloh, F., Saak, W, Haase, D. and Pohl, S. (1997) Synthesis, X-ray structure and electrochemical characterization of a binuclear thiolate bridged Ni-Fe-nitrosyl complex, related to the active site of NiFe hydrogenase. Chem. Commun. (Cambridge), 1997, 979-80. 

Rakhely, G., Zhou, Z. FI., Adams, M. W. and Kovacs, K. L. (1999) Biochemical and molecular characterization of the [NiEe] hydrogenase from the hyperthermophilic archaeon. Thermococcus litoralis. Eur. J. Biochem., 266, 1158-65. 

Rossmann, R., Maier, T., Lottspeich, F. and Bock, A. (1995) Characterization of a protease from Escherichia coli involved in hydrogenase maturation, Eur. J. Biochem., 227, 545-50. 

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