title: The 15 N-terminal amino acids of hexokinase II are not required for in vivo function: analysis of a truncated form of hexokinase II in Saccharomyces cerevisiae.
abstract: The function of the N-terminal amino acids of Saccharomyces cerevisiae hexokinase II was studied in vivo using strains producing a form of hexokinase II lacking its first 15 amino acids (short form). This short form of hexokinase II was produced from a fusion between the promoter region of the PGK1 gene and the HXK2 coding sequence except the first 15 codons. As expected, the in vitro analysis of the short form protein by gel filtration chromatography indicates that the short protein does not form dimers under conditions where the wild-type protein dimerizes. Kinetic studies show that the enzymatic activities are very similar to wild-type behavior. The physiological experiments performed on the strains containing the fusion allele demonstrate that the short form of the enzyme is similar to the wild-type both in terms of phosphorylation of hexoses and glucose repression. We conclude that the N-terminal amino acids of hexokinase II are not required in vivo either for phosphorylation of hexoses or for glucose repression.
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
authors.forename: H
authors.initials: H
authors.lastname: Ma
authors.forename: L M
authors.initials: LM
authors.lastname: Bloom
authors.forename: S E
authors.initials: SE
authors.lastname: Dakin
authors.forename: C T
authors.initials: CT
authors.lastname: Walsh
authors.forename: D
authors.initials: D
authors.lastname: Botstein
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D015153:Blotting, Western; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D003062:Codon; D004591:Electrophoresis, Polyacrylamide Gel; D006593:Hexokinase / Q000032:analysis / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D014170:Transformation, Genetic
chemical_list: D003062:Codon; D006593:Hexokinase