title: Yeast acyl-CoA-binding protein: acyl-CoA-binding affinity and effect on intracellular acyl-CoA pool size.
abstract: Acyl-CoA-binding protein (ACBP) is a 10 kDa protein characterized in vertebrates. We have isolated two ACBP homologues from the yeast Saccharomyces carlsbergensis, named yeast ACBP types 1 and 2. Both proteins contain 86 amino acid residues and are identical except for four conservative substitutions. In comparison with human ACBP, yeast ACBPs exhibit 48% (type 1) and 49% (type 2) conservation of amino acid residues. The amino acid sequence of S. carlsbergensis ACBP type 1 was found to be identical with the one ACBP present in Saccharomyces cerevisiae. A recombinant form of this protein was expressed in Escherichia coli and S. cerevisiae, purified, and its acyl-CoA-binding properties were characterized by isoelectric focusing and microcalorimetric analyses. The yeast ACBP was found to bind acyl-CoA esters with high affinity (Kd 0.55 x 10(-10) M). Overexpression of yeast ACBP in S. cerevisiae resulted in a significant expansion of the intracellular acyl-CoA pool. Finally, Southern-blotting analysis of the two genes encoding ACBP types 1 and 2 in S. carlsbergensis strongly indicated that this species is a hybrid between S. cerevisiae and Saccharomyces monacensis.
authors.affiliation: Institute of Biochemistry, Odense University, Denmark.
authors.forename: J
authors.initials: J
authors.lastname: Knudsen
authors.forename: N J
authors.initials: NJ
authors.lastname: Faergeman
authors.forename: H
authors.initials: H
authors.lastname: Skøtt
authors.forename: R
authors.initials: R
authors.lastname: Hummel
authors.forename: C
authors.initials: C
authors.lastname: Børsting
authors.forename: T M
authors.initials: TM
authors.lastname: Rose
authors.forename: J S
authors.initials: JS
authors.lastname: Andersen
authors.forename: P
authors.initials: P
authors.lastname: Højrup
authors.forename: P
authors.initials: P
authors.lastname: Roepstorff
authors.forename: K
authors.initials: K
authors.lastname: Kristiansen
mesh_terms: D000214:Acyl Coenzyme A / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D002151:Calorimetry; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D026261:Diazepam Binding Inhibitor; D004578:Electron Spin Resonance Spectroscopy; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005800:Genes, Fungal; D016681:Genome, Fungal; D006801:Humans; D007526:Isoelectric Point; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012440:Saccharomyces / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D000214:Acyl Coenzyme A; D002352:Carrier Proteins; D026261:Diazepam Binding Inhibitor; D011994:Recombinant Proteins