a genetic and pharmacological analysis of isoprenoid pathway by lc-msms in fission yeast类异戊二烯途径的基因和药物分析lc-msms裂殖酵母.pdfVIP

A Genetic and Pharmacological Analysis of Isoprenoid Pathway by LC-MS/MS in Fission Yeast 1,2 1,3 1,4 1 1 1,3 Tomonori Takami , Yue Fang *, Xin Zhou , Wurentuya Jaiseng , Yan Ma , Takayoshi Kuno 1 Division of Molecular Pharmacology and Pharmacogenomics, Department of Biochemistry and Molecular Biology, Kobe University Graduate School of Medicine, Kobe, Japan, 2 Chemical Analysis Section, JCL Bioassay Corporation, Nishiwaki, Japan, 3 Department of Pharmacology, School of Pharmaceutical Sciences, China Medical University, Shenyang, China, 4 The First Affiliated Hospital of Liaoning Medical University, Jinzhou, China Abstract Currently, statins are the only drugs acting on the mammalian isoprenoid pathway. The mammalian genes in this pathway are not easily amenable to genetic manipulation. Thus, it is difficult to study the effects of the inhibition of various enzymes on the intermediate and final products in the isoprenoid pathway. In fission yeast, antifungal compounds such as azoles and terbinafine are available as inhibitors of the pathway in addition to statins, and various isoprenoid pathway mutants are also available. Here in these mutants, treated with statins or antifungals, we quantified the final and intermediate products of the fission yeast isoprenoid pathway using liquid chromatography-mass spectrometry/mass spectrometry. In hmg1-1, a mutant of the gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR), ergosterol (a final sterol product), and squalene (an intermediate pathway product), were decreased to approximately 80% and 10%, respectively, compared with that of wild-type cells. Consistently in wild-type cells, pravastatin, an HMGR inhibitor decreased ergosterol and squalene, and