title: A novel copper-binding protein with characteristics of a metallothionein from a clinical isolate of Candida albicans.
abstract: It is known that clinical isolates of Candida albicans exhibit a high level of resistance to copper salts, although the molecular basis of this resistance is not clear. To investigate this, a novel copper-binding protein was purified from a clinical isolate of C. albicans. The protein was extracted from yeast cells after an induction period of 10 h in a copper-containing suspension medium. It was further purified by size-exclusion chromatography, ultrafiltration and reverse-phase HPLC. All protein fractions were analysed for their protein and copper contents. The copper/protein ratio increased steadily throughout the purification process; the most highly purified fraction showed a 210-fold increase compared to the whole-cell extract, with a recovery of 0.03%. The molecular mass of the protein was 10,000 Da and a reconstitution study using the purified apoprotein suggested that the equivalent extent of Cu(I) binding was approximately 14 mol eq. The amino-terminal segment of the copper-binding protein revealed three Cys-Xaa-Cys motifs, which is typical of a metallothionein (MT), and showed significant homology with mammalian MTs with respect to the positions of the cysteine residues. This is the first report of the isolation of a copper-binding protein from C. albicans.
authors.affiliation: Department of Biotechnology, Faculty of Technology, Tokyo University of Agriculture and Technology, 2-24-16 Nakamachi, Koganei, Tokyo 184-8588, Japan1.
authors.forename: Ki-Bong
authors.initials: KB
authors.lastname: Oh
authors.affiliation: Department of Biotechnology, Faculty of Technology, Tokyo University of Agriculture and Technology, 2-24-16 Nakamachi, Koganei, Tokyo 184-8588, Japan1.
authors.forename: Takahide
authors.initials: T
authors.lastname: Watanabe
authors.affiliation: Department of Biotechnology, Faculty of Technology, Tokyo University of Agriculture and Technology, 2-24-16 Nakamachi, Koganei, Tokyo 184-8588, Japan1.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Matsuoka
mesh_terms: D000595:Amino Acid Sequence; D001059:Apoproteins / Q000737:chemistry / Q000302:isolation & purification; D002176:Candida albicans / Q000737:chemistry* / Q000302:isolation & purification; D002177:Candidiasis / Q000382:microbiology*; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D003300:Copper / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008668:Metallothionein / Q000378:metabolism; D008969:Molecular Sequence Data
chemical_list: D001059:Apoproteins; D002352:Carrier Proteins; C023548:copper-binding protein; D003300:Copper; D008668:Metallothionein