title: Steady-state kinetics of thiocyanate oxidation catalyzed by human salivary peroxidase.
abstract: A steady-state kinetic analysis was made of thiocyanate (SCN-) oxidation catalyzed by human peroxidase (SPO) isolated from parotid saliva. For comparative purposes, bovine lactoperoxidase (LPO) was also studied. Both enzymes followed the classical Theorell-Chance mechanism under the initial conditions [H2O2] less than 0.2mM, [SCN-] less than 10mM, and pH greater than 6.0. The pH-independent rate constants (k1) for the formation of compound I were estimated to be 8 X 10(6) M-1 s-1 (SD = 1, n = 18) for LPO and 5 X 10(6) M-1 s-1 (SD = 1, n = 11) for SPO. The pH-independent second-order rate constants (k4) for the oxidation of thiocyanate by compound I were estimated to be 5 X 10(6) M-1 s-1 (SD = 1, n = 18) for LPO and 9 X 10(6) M-1 s-1 (SD = 2, n = 11) for SPO. Both enzymes were inhibited by SCN- at pH less than 6. The pH-independent equilibrium constant (Ki) for the formation of the inhibited enzyme-SCN- complex was estimated to be 24 M-1 (SD = 12, n = 8) for LPO and 44 M-1 (SD = 4, n = 10) for SPO. An apparent pH dependence of the estimated values for k4 and Ki for both LPO and SPO was consistent with a mechanism based on assumptions that protonation of compound I was necessary for the SCN- peroxidation step, that a second protonation of compound I gave an inactive form, and that the inhibited enzyme-SCN- complex could be further protonated to give another inactive form.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Biochemistry, University of Alabama, Birmingham 35294.
authors.forename: K M
authors.initials: KM
authors.lastname: Pruitt
authors.forename: B
authors.initials: B
authors.lastname: Mansson-Rahemtulla
authors.forename: D C
authors.initials: DC
authors.lastname: Baldone
authors.forename: F
authors.initials: F
authors.lastname: Rahemtulla
mesh_terms: D006801:Humans; D007700:Kinetics; D010084:Oxidation-Reduction; D010306:Parotid Gland / Q000201:enzymology; D010544:Peroxidases / Q000378:metabolism*; D012463:Saliva / Q000201:enzymology*; D013861:Thiocyanates / Q000378:metabolism*
chemical_list: D013861:Thiocyanates; D010544:Peroxidases