Chemoselective Attachment of Biologically Active Proteins to Surfaces by Expressed Protein Ligation and Its Application for “Protein Chip” Fabrication
- 23 October 2004
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 126 (45) , 14730-14731
- https://doi.org/10.1021/ja0456611
Abstract
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.Keywords
This publication has 15 references indexed in Scilit:
- The Staudinger Ligation—A Gift to Chemical BiologyAngewandte Chemie International Edition in English, 2004
- Electric Current through a Molecular Rod—Relevance of the Position of the Anchor GroupsAngewandte Chemie International Edition in English, 2003
- Site-Specific Protein Immobilization by Staudinger LigationJournal of the American Chemical Society, 2003
- Protein Semi-Synthesis in Living CellsJournal of the American Chemical Society, 2003
- Protein analysis on a proteomic scaleNature, 2003
- Niobium-Zirconium Chronometry and Early Solar System DevelopmentScience, 2002
- Synthesis of Native Proteins by Chemical LigationAnnual Review of Biochemistry, 2000
- Studying Receptor−Ligand Interactions Using Encoded Amino Acid ScanningBiochemistry, 1998
- Synthesis of Proteins by Native Chemical LigationScience, 1994
- Constructing Proteins by Dovetailing Unprotected Synthetic Peptides: Backbone-Engineered HIV ProteaseScience, 1992