Nickel-deficient carbon monoxide dehydrogenase from Rhodospirillum rubrum: in vivo and in vitro activation by exogenous nickel
An inactive, Ni-deficient form of carbon monoxide (CO) dehydrogenase designated apo-CO dehydrogenase, accumulated in Rhodospirillum rubrum when cells were grown in the absence of Ni and treated with CO. In vivo, both CO dehydrogenase activity and hydrogenase activity increased several hundred fold upon addition of 2 ..mu..M NiCl/sub 2/. Apo-CO dehydrogenase was purified to homogeneity and differed from holo-CO dehydrogenase only in its activity and Ni content, containing <0.2 mol of Ni per mol of protein, and a specific activity of 35 ..mu..mol of CO per min per mg. Optimal in vitro activation of purified apo-CO hydrogenase resulted in an enzyme with a specific activity of 2640 ..mu..mol of CO per min per mg. No additional enzymes or low molecular weight cofactors were required for activation. Apo-CO dehydrogenase was not activated by MgCl/sub 2/, MnCl/sub 2/, CuCl/sub 2/, ZnCl/sub 2/, CoCl/sub 2/, or Na/sub 2/MoO/sub 4/. /sup 63/Ni was incorporated into apo-CO dehydrogenase during activation. The electron paramagnetic resonance (EPR) spectra of dithionite-reduced apo- and holo-enzyme were identical, indicating that, in the reduced state, the Fe-S centers observed by EPR are unchanged in the apo-enzyme.
- Research Organization:
- Univ. of Wisconsin, Madison (USA)
- OSTI ID:
- 6470067
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 85:1
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
NICKEL COMPOUNDS
BIOLOGICAL EFFECTS
OXIDOREDUCTASES
ENZYME INDUCTION
RHODOSPIRILLUM
ENZYME ACTIVITY
CARBON MONOXIDE
ELECTRON SPIN RESONANCE
ELECTROPHORESIS
IN VITRO
IN VIVO
MOLECULAR STRUCTURE
NICKEL 63
BACTERIA
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBON COMPOUNDS
CARBON OXIDES
CHALCOGENIDES
ENZYMES
EVEN-ODD NUCLEI
GENE REGULATION
INTERMEDIATE MASS NUCLEI
ISOTOPES
MAGNETIC RESONANCE
MICROORGANISMS
NICKEL ISOTOPES
NUCLEI
OXIDES
OXYGEN COMPOUNDS
RADIOISOTOPES
RESONANCE
TRANSITION ELEMENT COMPOUNDS
YEARS LIVING RADIOISOTOPES
550201* - Biochemistry- Tracer Techniques