Intein-mediated generation of N-terminal cysteine proteins and their applications in live cell bioimaging and protein microarray

YEO SU-YIN, DAWN

Publication

Intein-mediated generation of N-terminal cysteine proteins and their applications in live cell bioimaging and protein microarray

YEO SU-YIN, DAWN

Abstract

The post-genomic era heralds a multitude of challenges for chemists and biologists alike, with the study of protein functions at the heart of much research. The elucidation of protein structure, localization, stability, post-translational modifications and protein interactions will steadily unveil the role of each protein and its associated biological function in the cell. Combining recombinant DNA technology, protein splicing, organic chemistry and the unique chemistry of native chemical ligation, we have developed a strategy to chemoselectively label proteins, both in vitro and in live cells, with biotin, fluorescent and other molecular probes. The site-specific incorporation of molecular entities with interesting chemical functionalities in proteins has many potential applications in chemical and biological studies of proteins. In this study, we present an intein-mediated approach to generate N-terminal cysteine containing proteins and its applications in two different areas related to proteomics and chemical biology, namely protein microarray technologies for large-scale protein analysis and live cell bioimaging.