title: A comparison of Zn(II) and Co(II) in the kinetics of inactivation of aminoacylase by 1,10-phenanthroline and reconstitution of the apoenzyme.
abstract: The kinetics of reconstitution of apoacylase with either Zn(II) or Co(II) and the inactivation of the Co(II) reconstituted enzyme by 1,10-phenanthroline (OP) has been studied by following the substrate reaction continuously in presence of the metal ion or OP respectively. Although the native Zn(II)-containing and the Co(II)-reconstituted enzymes have closely similar Michaelis constants and maximal velocities, the kinetics for both the inactivation by OP and the reconstitution of the apoenzyme with the metal ions differs considerably. For Co(II), both the inactivation by OP and the reconstitution show simple kinetics, but for Zn(II), the inhibition by OP is a multi-phasic process [Wang, Wu, Wang, Zhou and Tsou (1992) Biochem. J. 281, 285-290], and the kinetics of reconstitution is also much more complicated. Both the native and the Co(II)-reconstituted enzymes are inhibited by excess of Zn(II), but not by Co(II). The inhibition by Zn(II) in excess and the reconstitution of the apoenzyme with Zn(II) are co-operative processes. The inhibition by Zn and its effect on the fluorescence emission of 1-anilinonaphthalene-8-sulphonic acid bound to the native enzyme indicate multiple Zn(II)-binding sites.
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Academia Sinica, Beijing, China.
authors.forename: H B
authors.initials: HB
authors.lastname: Wu
authors.forename: C L
authors.initials: CL
authors.lastname: Tsou
mesh_terms: D000581:Amidohydrolases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D001051:Apoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D003035:Cobalt / Q000494:pharmacology*; D007700:Kinetics; D008433:Mathematics; D008962:Models, Theoretical; D010618:Phenanthrolines / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence; D015032:Zinc / Q000494:pharmacology*
chemical_list: D001051:Apoenzymes; D010618:Phenanthrolines; D003035:Cobalt; D000581:Amidohydrolases; C019672:aminoacylase I; D015032:Zinc; C025205:1,10-phenanthroline