Camarero Julio A, Kwon Youngeun, Coleman Matthew A
Chemical Biology and Nuclear Sciences Division and Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, 7000 East Avenue, Livermore, CA 94550, USA.
J Am Chem Soc. 2004 Nov 17;126(45):14730-1. doi: 10.1021/ja0456611.
The present work describes a general method for the selective attachment of proteins to solid surfaces through their C-termini that can be used for the efficient creation of protein chips. Our method is based in the chemoselective reaction between a protein C-terminal alpha-thioester and a modified surface containing N-terminal Cys residues. alpha-Thioester proteins can be obtained using standard recombinant techniques by using expression vectors containing engineered inteins. This new method was used to immobilize two fluorescent proteins and a functional SH3 domain using a protein microarrayer.
本研究描述了一种通过蛋白质C末端将蛋白质选择性连接到固体表面的通用方法，该方法可用于高效制备蛋白质芯片。我们的方法基于蛋白质C末端α-硫酯与含有N末端半胱氨酸残基的修饰表面之间的化学选择性反应。α-硫酯蛋白可通过使用含有工程化内含肽的表达载体，利用标准重组技术获得。利用蛋白质微阵列仪，采用这种新方法固定了两种荧光蛋白和一个功能性SH3结构域。
