Molecular cloning and biochemical characterization of Candida albicans acyl-CoA:sterol acyltransferase, a potential target of antifungal agents

Ki Young Kim, Yu Kyong Shin, Jong Chul Park, Jung Ho Kim, Hongyuan Yang, Dong Min Han, Young Ki Paik

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)

Abstract

To determine whether Candida albicans acyl CoA:sterol acyltransferase (ASAT) can be a potential target enzyme for the protoberberine derivative (HWY-289), we have isolated a gene encoding Ca-ASAT and examined inhibitory effects of HWY-289 on the overexpressed Ca-ASAT. HWY-289 specifically inhibits Ca-ASAT in a non-competitive manner in vitro (IC50 [9.2μM], K i [5.15μM]). The cloned CaARE2 gene (1830 nucleotides [nt]) encodes active Ca-ASAT protein that exhibits a calculated molecular mass of 71.3kDa. The amino acid sequence of CaAre2p is 33.4% and 35.1% identical to those of Saccharomyces cerevisiae ScAre1p and ScAre2p homologues, respectively. Recombinant and endogenous Ca-ASAT displayed identical patterns of inhibition upon exposure to HWY-289 and a preference for cholesterol and oleoyl-CoA as substrates. Northern blot analysis showed that CaARE2 was activated by HWY-289, but not by CI-976 (a human acyl-coenzyme A:cholesterol acyltransferase inhibitor), in a dose-dependent manner (up to 5mg/L), suggesting different selectivities of action between HWY-289 and CI-976 on Ca-ASAT activity.

Original languageEnglish
Pages (from-to)911-919
Number of pages9
JournalBiochemical and Biophysical Research Communications
Volume319
Issue number3
DOIs
Publication statusPublished - 2004 Jul 2

Bibliographical note

Funding Information:
We thank Drs. Catherine Chang and Ta Yuan Chang at Dartmouth College for their generosity in supplying CI-976, an ACAT inhibitor. We also thank Dr. Stephen L. Sturley (Columbia, NY, USA) for provision of sterol ester deficient yeast and Dr. Chery A. Gale (Minesota, USA) for provision of pMG1602. This study was supported by a grant of the Korea Health 21 R&D Project, Ministry of Health and Welfare, Republic of Korea (03-PJ10-PG6-GP01-002 toY.K.P.), Seoul, Korea.

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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