Diverse functionalization of Aurora-A kinase at specified surface and buried sites by native chemical modification.

ABSTRACT: The ability to obtain a homogeneous sample of protein is invaluable when studying the effect of alterations such as post-translational modifications (PTMs). Selective functionalization of a protein to investigate the effect of PTMs on its structure or activity can be achieved by chemical modification of cysteine residues. We demonstrate here that one such technique, which involves conversion of cysteine to dehydroalanine followed by thiol nucleophile addition, is suitable for the site-specific installation of a wide range of chemical mimics of PTMs, including acetylated and dimethylated lysine, and other unnatural amino acids. These reactions, optimized for the clinically relevant kinase Aurora-A, readily proceed to completion as revealed by intact protein mass spectrometry. Moreover, thes

SUBMITTER: Rowan F 

PROVIDER: S-EPMC4122486 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature