【2017年整理】过氧化氢酶3-氨基-1,2,4-三唑(ATM)的抑制剂在酿酒酵母中过氧化氢酶的生理作用.docVIP

Inhibition of Catalase by Aminotriazole in vivo Results in Reduction of Glucose-6-phosphate Dehydrogenase Activity in Saccharomyces cerevisiae CellsM. Bayliak, D. Gospodaryov, H. Semchyshyn, and V. LushchakDepartment of Biochemistry, Vassyl Stefanyk Precarpathian National University, 57 Shevchenko Str.,Ivano
Frankivsk 76025, Ukraine; fax: +38
0342
714683; E
mail: lushchak@pu.if.uaReceived August 3, 2007Revision received November 19, 2007 Abstract—The inhibitor of catalase 3-amino-1,2,4-triazole (AMT) was used to study the physiological role of catalase in the yeast Saccharomyces cerevisiae under starvation. It was shown that AMT at the concentration of 10 mM did not affect the growth of the yeast. In vivo and in vitro the degree of catalase inhibition by AMT was concentration
and time
dependent. Peroxisomal catalase in bakers’ yeast was more sensitive to AMT than the cytosolic one. In vivo inhibition of catalase byAMT in S. cerevisiae caused a simultaneous decrease in glucose-6-phosphate dehydrogenase activity and an increase in glutathione reductase activity. At the same time, the level of protein carbonyls, a marker of oxidative modification, was not affected. Possible mechanisms compensating the negative effects caused by AMT inhibition of catalase are discussed.摘要 过氧化氢酶3-氨基-1,2,4-三唑（ATM）的抑制剂常用来研究饥饿情况下酿酒酵母中过氧化氢酶的生理作用。结果表明ATM浓度在10 mM时，不会影响酵母的生长。在体内外过氧化氢酶的抑制程度由ATM的浓度和试剂有关。面包酵母中的过氧化氢酶对ATM比细胞质基质中的敏感。酿酒酵母中ATM对体内过氧化氢酶的抑制作用会同时降低葡萄糖-6-磷酸脱氢酶活性，同时增加谷胱甘肽还原酶的活性。同时，蛋白质金属羰基合物的水平（氧化修饰的一个标志）不受影响。补偿负效应的机制可能是由AMT抑制过氧化氢酶引起的。材料和方法化学药品和酵母菌株 葡萄糖-6-磷酸脱氢酶、苯甲基磺酰氟（PMSF）、异柠檬酸、EDTA和ATM都是从Sigma (USA) 获得的；NADP(H)、NAD(H)、2,4-二硝基苯肼、槲黄素、氧化型谷胱甘肽和TEMED是从Reanal (Hungary) 购买的，胍-HCl是从Fluka (Germany) 买的。无机化学材料是从Reakhim (Russia) 获得的。其他所有的化学药品都是高纯度的。酵母提取物是从BioGene (Great Britain) 获得的。本实验中使用的酿酒酵母菌株如下：YPH250 (MATa trp1-Δ1 his3-Δ200 lys2-801 leu2-Δ1 ade2-101 ura3-52)、YTT7 (YPH250 Δctt1::URA3)、YIT2 (YPH250 Δcta1::TRP1) 和 YWT1 (YPH250 Δcta1::TRP1 Δctt1::URA3)。这些菌株都是由Professor Yoshiharu Inoue (Kyoto Univers