Caplin B E, Marshall M S
Department of Medicine, Indiana University, Indianapolis 46202, USA.
Methods Enzymol. 1995;250:51-68. doi: 10.1016/0076-6879(95)50062-6.
The use of the S. cerevisiae protein prenyltransferases as a model system for general prenyltransferase study is justified by the similarity of mechanism, substrate specificity, and evolutionarily conserved substrates with the mammalian prenyltransferases. Genetic identification of potential structural genes involved in prenyltransferase activity can be easily confirmed with biochemical assays using recombinant enzyme reconstitution. Yeast FTase and GGTase I produced in E. coli are indistinguishable from the native proteins and can be studied without interference from contaminating cellular protein prenyltransferases. Structure-function analysis of the yeast prenyltransferase subunits is also simplified by the rapidity with which mutant enzymes can be analyzed in E. coli and their biological activity characterized in yeast defective for the particular subunit gene.
将酿酒酵母蛋白异戊二烯基转移酶用作一般异戊二烯基转移酶研究的模型系统是合理的，因为其机制、底物特异性以及与哺乳动物异戊二烯基转移酶在进化上保守的底物具有相似性。通过使用重组酶重组的生化测定，可以轻松确认参与异戊二烯基转移酶活性的潜在结构基因的遗传鉴定。在大肠杆菌中产生的酵母法尼基转移酶（FTase）和γ-谷氨酰转移酶I（GGTase I）与天然蛋白质无法区分，并且可以在不受污染的细胞蛋白异戊二烯基转移酶干扰的情况下进行研究。酵母异戊二烯基转移酶亚基的结构-功能分析也因在大肠杆菌中可以快速分析突变酶并在特定亚基基因缺陷的酵母中表征其生物活性而得以简化。
