Archives of Microbiology

, Volume 140, Issue 2–3, pp 206–211 | Cite as

Nickel and iron incorporation into soluble hydrogenase of Alcaligenes eutrophus

  • Cornelius G. Friedrich
  • Sergej Suetin
  • Michael Lohmeyer
Original Papers

Abstract

Qualitative and quantitative determination of proteins of the soluble hydrogenase (hydrogen: NAD+ oxidoreductase, EC 1.12.1.2) from Alcaligenes eutrophus H16 was done by crossed and rocket immunoelectrophoresis. Under metal sufficient growth conditions active hydrogenase (mol. wt. 200,000) cross reacting material (crmI) was formed. Under nickel limitation crmIII hydrogenase proteins were formed exhibiting diaphorase activity with a mol. wt. of 92,000±5,000. Other immunochemically active hydrogenase proteins present in significant amounts were not detected in the absence of nickel. Nickel-or ferric chloride induced active hydrogenase synthesis. Incorporation of both metals into the enzyme required protein synthesis as evident from studies inhibiting translation by chloramphenicol. Iron is bound to acid-labile sulfur in the hydrogenases. Since nickel incorporation was identical to that of iron, linkage of nickel to the soluble hydrogenase may be similar to that of iron. Studies inhibiting transcription suggested an unusual long half life of the hydrogenase mRNA of 1.5h.

Key words

Alcaligenes eutrophus Nickel in hydrogenase Nickel-free hydrogenase Immunochemistry of hydrogenase proteins mRNA half life 

Abbreviations

CIE

crossed immunoelectrophoresis

RIE

rocket immunoelectrophoresis

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Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • Cornelius G. Friedrich
    • 1
  • Sergej Suetin
    • 1
  • Michael Lohmeyer
    • 1
  1. 1.Institut für Mikrobiologie der Universität GöttingenGöttingenFederal Republic of Germany

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