title: Kinetics of complexing activation by the magnesium ion on green crab (Scylla serrata) alkaline phosphatase.
abstract: As with mammalian enzymes, green crab (Scylla serrata) alkaline phosphatase can be activated by Mg2+ through a time-dependent course. The activation is mainly a Vmax effect. Tsou's method was used to study the kinetic course of activation. The results show that the enzyme was activated by a complexing scheme that had not been previously identified: the enzyme first reversibly and quickly binds Mg2+ and then undergoes a slow reversible course to activation, with a relatively high activation energy (78 +/- 4 kJ/mol) and a slow conformational change. The activation reaction is a single molecule reaction, and the apparent activation rate constant is independent of Mg2+ concentration if the concentration is sufficiently high. The microscopic rate constants of activation and the association constant were determined from the measurements. The proposed scheme may also be applied to the Mg2+ activation mechanism for mammalian enzyme, to explain why the activation rate is time-dependent and not diffusion controlled. Substrate binding was also shown to affect the activation rate constant.
authors.affiliation: Department of Biological Science and Biotechnology, Tsinghua University, Beijing, People's Republic of China.
authors.forename: Y D
authors.initials: YD
authors.lastname: Park
authors.forename: Y
authors.initials: Y
authors.lastname: Yang
authors.forename: Q X
authors.initials: QX
authors.lastname: Chen
authors.forename: H N
authors.initials: HN
authors.lastname: Lin
authors.forename: Q
authors.initials: Q
authors.lastname: Liu
authors.forename: H M
authors.initials: HM
authors.lastname: Zhou
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D003386:Brachyura / Q000201:enzymology* / Q000502:physiology; D004789:Enzyme Activation; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013696:Temperature
chemical_list: D000469:Alkaline Phosphatase; D008274:Magnesium