Clark Kevin M, van der Donk Wilfred A, Lu Yi
Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
Methods Enzymol. 2009;462:97-115. doi: 10.1016/S0076-6879(09)62005-X.
Metalloproteins contain highly specialized metal-binding sites that are designed to accept specific metal ions to maintain correct function. Although many of the sites have been modified with success, the relative paucity of functional group availability within proteinogenic amino acids can sometimes leave open questions about specific functions of the metal binding ligands. Attaining a more thorough analysis of individual amino acid function within metalloproteins has been realized using expressed protein ligation (EPL). Here we describe our recent efforts using EPL to incorporate nonproteinogenic cysteine and methionine analogues into the type 1 copper site found in Pseudomonas aeruginosa azurin.
金属蛋白含有高度专业化的金属结合位点，这些位点旨在接受特定的金属离子以维持正确的功能。尽管许多位点已成功修饰，但蛋白质氨基酸内可用官能团的相对稀缺有时会留下有关金属结合配体特定功能的疑问。使用表达蛋白连接（EPL）已实现对金属蛋白中单个氨基酸功能进行更全面的分析。在此，我们描述了我们最近使用EPL将非蛋白质半胱氨酸和蛋氨酸类似物掺入铜绿假单胞菌天青蛋白中发现的1型铜位点的工作。
