Chemoselective Attachment of Biologically Active Proteins to Surfaces by Expressed Protein Ligation and Its Application for “Protein Chip” Fabrication

Julio A. Camarero,*† Youngeun Kwon,† and Matthew A. Coleman‡
Chemical Biology and Nuclear Sciences Division and Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, 7000 East Avenue, Livermore, California 94550
J. Am. Chem. Soc., 2004, 126 (45), pp 14730–14731
DOI: 10.1021/ja0456611
Publication Date (Web): October 23, 2004
Copyright © 2004 American Chemical Society
*

In papers with more than one author, the asterisk indicates the name of the author to whom inquiries about the paper should be addressed.

,
†

 Chemical Biology and Nuclear Sciences Division, Lawrence Livermore National Laboratory.

,
‡

 Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory.

, camarero1@llnl.gov

Abstract

Abstract Image

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 α-thioester and a modified surface containing N-terminal Cys residues. α-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.

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History

  • Published In Issue November 17, 2004
  • Received July 19, 2004

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