Journal of Biological Chemistry · 1966 · 59 citations · 22 references
Life SciencesBiochemistryMolecular BiologyEnzyme MechanismEnzymologyProtein BiochemistryDisulfide Bond FormationEnzyme CharacterizationEnzyme IsolationEnzyme KineticsEnzyme PurificationGlutathione ConjugationDisulfide Bond ChemistryDisulfide Bond MispairingDisulfide ChemistryEquilibriumMichaelis ConstantMolecular Weight DeterminationPh DependenceReduced Glutathione
Abstract An enzyme that catalyzes the following sulfhydryl-disulfide interchange reaction has been purified 180-fold from bovine kidney. GSH + CoASSG ⇌ GSSG + CoA-SH In addition, it has been detected in most rat tissues studied. Its pH optimum is 8.2 and the equilibrium constant for the reaction is near unity at pH 6.9 and 25°. Michaelis constants for reduced glutathione and CoASSG are 3.3 x 10-4 m and 4.5 x 10-5 m, respectively. The molecular weight of the enzyme, determined by gel filtration, is approximately 12,000. Several unsymmetrical disulfides containing a glutathione residue are equally as effective as substrates. The enzyme, which becomes less active during storage, is largely reactivated by GSH. Its potential physiological significance is discussed.
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The Determination of Enzyme Dissociation Constants
Hans Lineweaver, Dean Burk · Journal of the American Chemical Society · 1934 · 12.1K citations
Devices for Graident Elution in Chromatography
Robert M. Bock, Nan-Sing Ling · Analytical Chemistry · 1954 · 328 citations