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Biochemistry. 1996 Apr 23;35(16):5345-52.

Metal ion interaction with urease and UreD-urease apoproteins.

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Department of Microbiology, Michigan State University, East Lansing 48824-1101, USA.


Klebsiella aerogenes urease in a Ni-containing enzyme (two Ni per alpha beta gamma unit) that is purified as an apoprotein from cells grown in Ni-free medium. Partial activation of urease and UreD-urease apoproteins is achieved in vitro by incubation in the presence of Ni(II) and CO2, whereas incubation of these proteins with Ni alone leads to the formation of inactive species [Park, I.-S., & Hausinger, R. P. (1995) Science 267, 1156-1158]. Here we determined the kinetics of these inhibitory reactions and demonstrated the presence of two Ni ions per alpha beta gamma unit in the inactive proteins. Although metal-substituted urease has never been purified from Ni-deprived cell, several other metal ions were shown to bind to the urease apoproteins. Divalent Zn, C, Co, and Mn all inhibited Ni- and Co2-promoted urease activation at concentrations below that of Ni, whereas Mg and Ca ions did not inhibit this process. Ni-inhibited species recovered their ability to be partially activated after EDTA treatment. In contrast, samples that were exposed to Co or Cu ions were irreversibly inactivated, and EDTA treatment of Zn- or Mn-inhibited samples led to reduced levels of activation competence. Mn-substituted urease, generated from urease apoprotein samples in a Mn- and Co2-dependent manner, was shown to be active, whereas other metal-substituted forms if urease lacked activity. The Mn-protein possessed only 2% of the activity of Ni-activated apoprotein [ approximately 8.0 vs approximately 400 mumol min-1 (mg protein)-1], but its KM value was only moderately altered from that of the native enzyme (3.86 +/- 0.15 mM vs 0.2 mM). Unlike the Ni-containing enzyme, Mn-urease was inhibited by EDTA. Given the evidence that urease apoprotein binds numerous metal ions, we speculate on possible roles for the UreD, UreF, and UreG accessory proteins in urease activation.

[Indexed for MEDLINE]

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