Kaplan J, DeGrado W F
Department of Biochemistry and Biophysics, School of Medicine, University of Pennsylvania, Philadelphia, PA 19104-6059, USA.
Proc Natl Acad Sci U S A. 2004 Aug 10;101(32):11566-70. doi: 10.1073/pnas.0404387101. Epub 2004 Aug 3.
The de novo design of catalytic proteins provides a stringent test of our understanding of enzyme function, while simultaneously laying the groundwork for the design of novel catalysts. Here we describe the design of an O(2)-dependent phenol oxidase whose structure, sequence, and activity are designed from first principles. The protein catalyzes the two-electron oxidation of 4-aminophenol (k(cat)/K(M) = 1,500 M(-1).min(-1)) to the corresponding quinone monoimine by using a diiron cofactor. The catalytic efficiency is sensitive to changes of the size of a methyl group in the protein, illustrating the specificity of the design.
催化蛋白的从头设计对我们对酶功能的理解提出了严格考验,同时也为新型催化剂的设计奠定了基础。在此,我们描述了一种依赖氧气的酚氧化酶的设计,其结构、序列和活性均基于第一原理进行设计。该蛋白利用双铁辅因子将4-氨基苯酚(k(cat)/K(M) = 1,500 M(-1)·min(-1))催化进行两电子氧化,生成相应的醌单亚胺。催化效率对蛋白中甲基大小的变化敏感,这说明了设计的特异性。