Identification of a bacterial energy-transducing ATPase as a metallo (Zn2+) protein. Effect of chelating agents and divalent metal ions on ATPase activity.
Purified F1-ATPase from Micrococcus lysodeikticus contains zinc in the amount of 1 mol/mol of enzyme. This zinc content correlates with standard values of ATPase activity (assayed with Ca2+-ATP as substrate) of the protein, i.e. 5--6 mumol substrate hydrolysed . min-1 . mg-1. Prolonged dialysis against EDTA results in a zinc-free protein which concomitantly loses its ATPase activity. Chelators such as Zincon, EDTA and L-cysteine inhibit the ATPase activity in concentration and/or time dependence related to their affinity for the metal ion involved. Reconstitution of the metallo (Zn2+) protein is demonstrated by the incorporation to the zinc-free protein of 65Zn2+ in amount near the 1 mol/mol of enzyme. This incorporation was concomitant with the regain of ATPase activity. The inhibition by EDTA and Zincon is reversed specifically by Zn2+ while the inhibition by EDTA is prevented by Zn2+ and Mn2+ and to, a minor extent, by Cd2+. Zn2+ and Ca2+ ions are involved and are probably mandatory in the ATPase activity of M. lysodeikticus F1 but their roles appear to be different and not exchangeable. Other divalent metal ions inhibit the Ca2+-ATPase activity of the Zn2+ protein by the following decreasing order; Hg2+, Fe2+, Co2+, Cd2+, Mn2+, Mg2+. M. lysodeikticus F1-ATPase is thus identified as a metallo (zinc) protein, which requires additional divalent metal ions for ATP hydrolysis.
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D002413:Cations, Divalent / Q000494:pharmacology; D002614:Chelating Agents / Q000494:pharmacology*; D004735:Energy Transfer; D008667:Metalloproteins / Q000378:metabolism*; D008670:Metals / Q000494:pharmacology*; D008837:Micrococcus / Q000201:enzymology; D015032:Zinc / Q000378:metabolism*
D002413:Cations, Divalent; D002614:Chelating Agents; D008667:Metalloproteins; D008670:Metals; D000251:Adenosine Triphosphatases; D015032:Zinc