Villegas V, Viguera A R, Avilés F X, Serrano L
Departament de Bioqulmica, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Fold Des. 1996;1(1):29-34.
Increasing protein stability is a major goal of protein engineering because of its potential industrial and pharmacological applications. Several different rule-of-thumb strategies have been employed for such a purpose, but a general rational method is still lacking. Recently, there has been significant progress in our understanding of the interactions responsible for helix stability in monomeric peptides and this information has been included in algorithms based on the helix/coil transition theory. We set out to investigate whether it is possible to use these algorithms to rationally increase protein stability.
Using a helix/coil transition algorithm, AGADIRms, we have designed mutations affecting solvent-exposed residues which, as predicted, significantly increase the helical stability in aqueous solution of peptides corresponding to the two alpha-helices of the activation domain of procarboxipeptidase A. Introduction of the same mutations in the protein results in proteins more resistant to urea or temperature denaturation, and there is a qualitative agreement between the expected and observed increases in stability.
In this work we demonstrate that by using a helix/coil algorithm to design helix-stabilizing mutations on the solvent-exposed face of helices, it is possible to rationally increase the stability of proteins.
由于蛋白质在工业和药理学方面的潜在应用，提高其稳定性是蛋白质工程的一个主要目标。为此已采用了几种不同的经验法则策略，但仍缺乏一种通用的合理方法。最近，我们对单体肽中负责螺旋稳定性的相互作用的理解取得了重大进展，并且这些信息已被纳入基于螺旋/卷曲转变理论的算法中。我们着手研究是否有可能使用这些算法来合理提高蛋白质的稳定性。
使用螺旋/卷曲转变算法AGADIRms，我们设计了影响溶剂暴露残基的突变，正如所预测的那样，这些突变显著提高了与羧肽酶A原激活域的两个α螺旋相对应的肽在水溶液中的螺旋稳定性。在蛋白质中引入相同的突变会使蛋白质对尿素或温度变性更具抗性，并且预期的稳定性增加与观察到的稳定性增加之间存在定性一致性。
在这项工作中，我们证明通过使用螺旋/卷曲算法在螺旋的溶剂暴露面上设计稳定螺旋的突变，可以合理提高蛋白质的稳定性。
