N-Myristoyl transferase-mediated protein labelling in vivo.
N-Myristoyl transferase-mediated labelling using a substrate modified with an azide or alkyne tag is described as an efficient and site-selective method for the introduction of a bioorthogonal tag at the N-terminus of a recombinant protein. The procedure may be performed in vitro, or in a single over-expression/tagging step in vivo in bacteria; tagged proteins may then be captured using Staudinger-Bertozzi or 'click' chemistry protocols to introduce a secondary label for downstream analysis. The straightforward synthesis of the chemical and molecular biological tools described should enable their use in a wide range of N-terminal labelling applications.
Heal
William P
WP
Department of Chemistry, South Kensington Campus, Imperial College, London, UK SW7 2AZ.
Wickramasinghe
Sasala R
SR
Leatherbarrow
Robin J
RJ
Tate
Edward W
EW
D000217:Acyltransferases / Q000378:metabolism*; D000480:Alkynes / Q000737:chemistry; D000595:Amino Acid Sequence; D001386:Azides / Q000737:chemistry; D002176:Candida albicans / Q000201:enzymology*; D008969:Molecular Sequence Data; D019814:Myristic Acid / Q000737:chemistry / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods*; D013379:Substrate Specificity
D000480:Alkynes; D001386:Azides; D010455:Peptides; D011994:Recombinant Proteins; D019814:Myristic Acid; D000217:Acyltransferases; C050480:glycylpeptide N-tetradecanoyltransferase