title: Nickel-deficient carbon monoxide dehydrogenase from Rhodospirillum rubrum: in vivo and in vitro activation by exogenous nickel.
abstract: An inactive, Ni-deficient form of carbon monoxide (CO) dehydrogenase [carbon-monoxide:(acceptor) oxidoreductase; EC 1.2.99.2], 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 microM NiCl2. Apo-CO dehydrogenase was purified to homogeneity and differed from holo-CO dehydrogenase only in its activity and Ni content, containing less than 0.2 mol of Ni per mol of protein, and a specific activity of 35 mumol of CO per min per mg. Optimal in vitro activation of purified apo-CO dehydrogenase resulted in an enzyme with a specific activity of 2640 mumol 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 MgCl2, MnCl2, CuCl2, ZnCl2, CoCl2, or Na2MoO4. 63Ni 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.
authors.affiliation: Department of Biochemistry, College of Agricultural and Life Sciences, University of Wisconsin, Madison 53706.
authors.forename: D
authors.initials: D
authors.lastname: Bonam
authors.forename: M C
authors.initials: MC
authors.lastname: McKenna
authors.forename: P J
authors.initials: PJ
authors.lastname: Stephens
authors.forename: P W
authors.initials: PW
authors.lastname: Ludden
mesh_terms: D000445:Aldehyde Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D004789:Enzyme Activation; D006864:Hydrogenase / Q000378:metabolism; D007700:Kinetics; D009097:Multienzyme Complexes; D009532:Nickel / Q000378:metabolism* / Q000494:pharmacology; D012247:Rhodospirillum rubrum / Q000201:enzymology*
chemical_list: D009097:Multienzyme Complexes; C022838:nickel chloride; D009532:Nickel; D006864:Hydrogenase; D000445:Aldehyde Oxidoreductases; C019823:carbon monoxide dehydrogenase