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Volume 145, Issue 9

A novel copper-binding protein with characteristics of a metallothionein from a clinical isolate of Free

Abstract

It is known that clinical isolates of 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 . 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 003%. The molecular mass of the protein was 10000 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 .

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  • Accepted:
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  • Published Online:
Keyword(s): amino-terminal sequence , Candida albicans , CBB, Coomassie brilliant blue , copper-binding protein , metallothionein , MT, metallothionein , protein purification and characterization and SDTC, sodium diethyldithiocarbamate
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1999-09-01
2024-04-18
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A novel copper-binding protein with characteristics of a metallothionein from a clinical isolate of Candida albicans
Microbiology 145, 2423 (1999); https://doi.org/10.1099/00221287-145-9-2423
/content/journal/micro/10.1099/00221287-145-9-2423
/content/journal/micro/10.1099/00221287-145-9-2423
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