title: Comparison of inactivation and unfolding of green crab (Scylla serrata) alkaline phosphatase during denaturation by guanidinium chloride.
abstract: Green crab (Scylla serrata) alkaline phosphatase (EC 3.1.3.1) is a metalloenzyme, each active site in which contains a tight cluster of two zinc ions and one magnesium ion. Unfolding and inactivation of the enzyme during denaturation in guanidinium chloride (GuHCl) solutions of different concentrations have been compared. The kinetic theory of the substrate reaction during irreversible inhibition of enzyme activity previously described by Tsou [(1988), Adv. Enzymol. Related Areas Mol. Biol. 61, 381-436] has been applied to a study on the kinetics of the course of inactivation of the enzyme during denaturation by GuHCl. The rate constants of unfolding and inactivation have been determined. The results show that inactivation occurs before noticeable conformational change can be detected. It is suggested that the active site of green crab alkaline phosphatase containing multiple metal ions is also situated in a limited region of the enzyme molecule that is more fragile to denaturants than the protein as a whole.
authors.affiliation: Department of Biology, Xiamen University, China.
authors.forename: Q X
authors.initials: QX
authors.lastname: Chen
authors.forename: W
authors.initials: W
authors.lastname: Zhang
authors.forename: W Z
authors.initials: WZ
authors.lastname: Zheng
authors.forename: Z
authors.initials: Z
authors.lastname: Zhang
authors.forename: S X
authors.initials: SX
authors.lastname: Yan
authors.forename: T
authors.initials: T
authors.lastname: Zhang
authors.forename: H M
authors.initials: HM
authors.lastname: Zhou
mesh_terms: D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D003386:Brachyura / Q000201:enzymology*; D002942:Circular Dichroism; D019791:Guanidine; D006146:Guanidines / Q000494:pharmacology; D007700:Kinetics; D008667:Metalloproteins / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D013050:Spectrometry, Fluorescence
chemical_list: D006146:Guanidines; D008667:Metalloproteins; D000469:Alkaline Phosphatase; D019791:Guanidine