Induction, localization and metal content of hydrogenase in the green alga Chlamydomonas reinhardtii

The hydrogenase enzyme occurring in Chlamydomonas reinhardtii is induced by anaerobic adaptation of the cells. In aerobically growing cells, antibodies against the hydrogenase failed to detect either active or inactive enzyme. However, already 10 min after the onset of anaerobic adaptation, the prot...

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Veröffentlicht in:European journal of biochemistry 1994-06, Vol.222 (3), p.769-774
Hauptverfasser: HAPPE, Thomas, MOSLER, Britta, NABER, J. Dirk
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container_title European journal of biochemistry
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creator HAPPE, Thomas
MOSLER, Britta
NABER, J. Dirk
description The hydrogenase enzyme occurring in Chlamydomonas reinhardtii is induced by anaerobic adaptation of the cells. In aerobically growing cells, antibodies against the hydrogenase failed to detect either active or inactive enzyme. However, already 10 min after the onset of anaerobic adaptation, the protein could be detected. The maximal amount of enzyme was reached after 2–3 hours anaerobiosis. Addition of nickel or iron to the growth medium did not influence activity. In atomic absorption experiments, a Ni/Fe ratio of about 1:250 was measured. We, therefore, propose the hydrogenase from C. reinhardtii to be of the Fe‐only type. Adaptation in the presence of uncouplers of phosphorylation showed this process to be energy‐dependent. From protein synthesis inhibition experiments, it is concluded that the protein is synthesized on cytoplasmic ribosomes and, therefore, must be nuclear encoded. After isolation of intact chloroplasts from adapted cells, the active enzyme was shown, by Western‐blotting analysis, to be located in the chloroplasts.
doi_str_mv 10.1111/j.1432-1033.1994.tb18923.x
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subjects Adaptation, Physiological
Anaerobiosis
Animals
Blotting, Western
Chlamydomonas reinhardtii
Chlamydomonas reinhardtii - enzymology
Chlamydomonas reinhardtii - growth & development
Chloramphenicol - pharmacology
Chloroplasts - enzymology
Cycloheximide - pharmacology
Enzyme Induction - drug effects
Freshwater
Hydrogenase - analysis
Hydrogenase - biosynthesis
Hydrogenase - chemistry
Iron - analysis
Iron - pharmacology
Nickel - pharmacology
Phosphorylation
Ribosomes - enzymology
Spectrophotometry, Atomic
Uncoupling Agents - pharmacology
title Induction, localization and metal content of hydrogenase in the green alga Chlamydomonas reinhardtii
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