Dwyer Mary A, Looger Loren L, Hellinga Homme W
Department of Biochemistry, Duke University Medical Center, Durham, NC 27710, USA.
Science. 2004 Jun 25;304(5679):1967-71. doi: 10.1126/science.1098432.
Rational design of enzymes is a stringent test of our understanding of protein chemistry and has numerous potential applications. Here, we present and experimentally validate the computational design of enzyme activity in proteins of known structure. We have predicted mutations that introduce triose phosphate isomerase activity into ribose-binding protein, a receptor that normally lacks enzyme activity. The resulting designs contain 18 to 22 mutations, exhibit 10(5)- to 10(6)-fold rate enhancements over the uncatalyzed reaction, and are biologically active, in that they support the growth of Escherichia coli under gluconeogenic conditions. The inherent generality of the design method suggests that many enzymes can be designed by this approach.
酶的合理设计是对我们蛋白质化学理解的严格考验，并且有众多潜在应用。在此，我们展示并通过实验验证了已知结构蛋白质中酶活性的计算设计。我们预测了将磷酸丙糖异构酶活性引入核糖结合蛋白的突变，核糖结合蛋白是一种通常缺乏酶活性的受体。所得设计包含18至22个突变，与未催化反应相比，反应速率提高了10⁵至10⁶倍，并且具有生物活性，因为它们在糖异生条件下支持大肠杆菌的生长。该设计方法的内在通用性表明，许多酶都可以通过这种方法进行设计。
