Kaiser E T, Lawrence D S, Rokita S E
Annu Rev Biochem. 1985;54:565-95. doi: 10.1146/annurev.bi.54.070185.003025.
The chemical modification of enzymes has played and will continue to play an important role in probing the mechanism of enzyme activity. This technique can be utilized for identification of those individual amino acid residues responsible for the catalytic properties of the entire protein. In chemical modification experiments, changes in enzymatic specificity have been noted, but often not predicted. However, in recent years, rational approaches for the alteration of enzymatic properties have become feasible by means of site-specific mutagenesis and chemical methodology. In the first method, one amino acid can be replaced by a new one; in the second method, not only can new amino acid residues be introduced but also new catalytic entities (such as flavins) can be affixed to the protein molecule. Both methodologies are in their infancy, yet they represent a potentially powerful approach toward the design and synthesis of enzymes possessing new specificities.
酶的化学修饰在探索酶活性机制方面已经并将继续发挥重要作用。该技术可用于鉴定那些对整个蛋白质的催化特性起作用的单个氨基酸残基。在化学修饰实验中,人们已经注意到酶特异性的变化,但往往无法预测。然而,近年来,通过定点诱变和化学方法来改变酶性质的合理方法已经变得可行。在第一种方法中,一个氨基酸可以被另一个新的氨基酸取代;在第二种方法中,不仅可以引入新的氨基酸残基,还可以将新的催化实体(如黄素)附着到蛋白质分子上。这两种方法都尚处于起步阶段,但它们代表了一种设计和合成具有新特异性的酶的潜在有力方法。