Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1982 Oct;152(1):42–48. doi: 10.1128/jb.152.1.42-48.1982

Nickel in the catalytically active hydrogenase of Alcaligenes eutrophus.

C G Friedrich, K Schneider, B Friedrich
PMCID: PMC221372  PMID: 6749813

Abstract

Nickel is a constituent of soluble and particulate hydrogenase of Alcaligenes eutrophus. Incorporation of 63Ni2+ revealed that almost the total nickel taken up by the cells was bound to the protein. Chromatography of a crude extract on diethylaminoethyl cellulose demonstrated an association of 63Ni2+ with soluble and particulate hydrogenase, supported by further analysis like polyacrylamide gel electrophoresis. Unspecific binding of 63Ni2+ to the protein was excluded by comparison with a mutant extract free of hydrogenase protein. X-ray fluorescence analysis of the homogeneous soluble hydrogenase indicated the presence of 2 mol of nickel per mol of enzyme, whereas the amount of nickel determined by incorporation of 63Ni2+ was calculated to be approximately 1 mol/mol of enzyme. Cells grown under nickel limitation contained catalytically inactive, but serologically active, soluble and particulate hydrogenase. The immunochemical reactions were only partially identical with the enzyme from nickel-cultivated cells indicating a structural modification of the proteins in the absence of nickel. It is concluded that nickel is essential for the catalytic activity of hydrogenase and not involved as a regulatory component in the synthesis of this enzyme.

Full text

PDF
42

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Albracht S. P., Graf E. G., Thauer R. K. The EPR properties of nickel in hydrogenase from Methanobacterium. FEBS Lett. 1982 Apr 19;140(2):311–313. doi: 10.1016/0014-5793(82)80921-6. [DOI] [PubMed] [Google Scholar]
  2. BARTHA R., ORDAL E. J. NICKEL-DEPENDENT CHEMOLITHOTROPHIC GROWTH OF TWO HYDROGENOMONAS STRAINS. J Bacteriol. 1965 Apr;89:1015–1019. doi: 10.1128/jb.89.4.1015-1019.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Diekert G., Klee B., Thauer R. K. Nickel, a component of factor F430 from Methanobacterium thermoautotrophicum. Arch Microbiol. 1980 Jan;124(1):103–106. doi: 10.1007/BF00407036. [DOI] [PubMed] [Google Scholar]
  4. Dixon N. E., Gazzola C., Blakeley R. L., Zerner B. Metal ions in enzymes using ammonia or amides. Science. 1976 Mar 19;191(4232):1144–1150. doi: 10.1126/science.769157. [DOI] [PubMed] [Google Scholar]
  5. Drake H. L. Occurrence of nickel in carbon monoxide dehydrogenase from Clostridium pasteurianum and Clostridium thermoaceticum. J Bacteriol. 1982 Feb;149(2):561–566. doi: 10.1128/jb.149.2.561-566.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Friedrich B., Friedrich C. G., Schlegel H. G. Purification and properties of chorismate mutase-prephenate dehydratase and prephenate dehydrogenase from Alcaligenes eutrophus. J Bacteriol. 1976 May;126(2):712–722. doi: 10.1128/jb.126.2.712-722.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Friedrich B., Hogrefe C., Schlegel H. G. Naturally occurring genetic transfer of hydrogen-oxidizing ability between strains of Alcaligenes eutrophus. J Bacteriol. 1981 Jul;147(1):198–205. doi: 10.1128/jb.147.1.198-205.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Schink B., Schlegel H. G. Mutants of Alcaligenes eutrophus defective in autotrophic metabolism. Arch Microbiol. 1978 May 30;117(2):123–129. doi: 10.1007/BF00402299. [DOI] [PubMed] [Google Scholar]
  9. Schink B., Schlegel H. G. The membrane-bound hydrogenase of Alcaligenes eutrophus. I. Solubilization, purification, and biochemical properties. Biochim Biophys Acta. 1979 Apr 12;567(2):315–324. doi: 10.1016/0005-2744(79)90117-7. [DOI] [PubMed] [Google Scholar]
  10. Schneider K., Cammack R., Schlegel H. G., Hall D. O. The iron-sulphur centres of soluble hydrogenase from Alcaligenes eutrophus. Biochim Biophys Acta. 1979 Jun 19;578(2):445–461. doi: 10.1016/0005-2795(79)90175-2. [DOI] [PubMed] [Google Scholar]
  11. Schneider K., Schlegel H. G. Purification and properties of soluble hydrogenase from Alcaligenes eutrophus H 16. Biochim Biophys Acta. 1976 Nov 8;452(1):66–80. doi: 10.1016/0005-2744(76)90058-9. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES