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PURIFICATION AND CHARACTERIZATION OF OX BRAIN NAD + ‐DEPENDENT ISOCITRATE DEHYDROGENASE
Author(s) -
Willson Vivian J. C.,
Tipton K. F.
Publication year - 1979
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1979.tb05270.x
Subject(s) - isocitrate dehydrogenase , nad+ kinase , idh1 , enzyme , affinity chromatography , allosteric regulation , biochemistry , chemistry , dehydrogenase , substrate (aquarium) , agarose , chromatography , biology , mutant , ecology , gene
— The NAD + ‐dependent isocitrate dehydrogenase from ox brain has been purified about 130‐fold by a method involving affinity chromatography on an NAD + ‐derivative of agarose. The enzyme preparation is not homogeneous but it is free from contaminating enzyme activities that could interfere with kinetic studies. The kinetic properties of the enzyme did not appear to have been altered by the purification procedure involved. The initial velocity of the reaction showed a sigmoid dependence on the concentration of isocitrate, and ADP behaved as an allosteric activator. The kinetics with NAD + as the substrate were hyperbolic. The molecular weight of the purified enzyme was found to be 285,000 ± 25,000.