title: Purification of the Type II and Type III isozymes of rat hexokinase, expressed in yeast.
abstract: C-terminally His-tagged versions of the Type II and Type III isozymes of rat hexokinase were expressed in Pichia pastoris and Schizosaccharomyces pombe, respectively. Milligram amounts of the homogeneous isozymes were readily obtained in good yield by chromatography on Ni-NTA columns. The specific activities were 133 +/- 4 and 76 +/- 3 u/mg for the purified Type II and Type III isozymes, respectively. The K(m)'s for glucose and ATP were in good agreement with values in the literature for the isozymes isolated from mammalian tissues. The Type III isozyme exhibited substrate inhibition at elevated levels of glucose, as previously observed for this isozyme isolated from mammalian tissue sources.
authors.affiliation: Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, Michigan 48824-1319, USA.
authors.forename: Dexin
authors.initials: D
authors.lastname: Su
authors.forename: John E
authors.initials: JE
authors.lastname: Wilson
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D015870:Gene Expression; D006593:Hexokinase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D010843:Pichia / Q000235:genetics; D051381:Rats; D012568:Schizosaccharomyces / Q000235:genetics
chemical_list: D007527:Isoenzymes; D006593:Hexokinase