N-Myristoyl transferase-mediated protein labelling in vivo
Author(s) -
William P. Heal,
Sasala R. Wickramasinghe,
Robin J. Leatherbarrow,
Edward W. Tate
Publication year - 2008
Publication title -
organic and biomolecular chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 146
eISSN - 1477-0539
pISSN - 1477-0520
DOI - 10.1039/b803258k
Subject(s) - bioorthogonal chemistry , labelling , chemistry , protein tag , click chemistry , transferase , biochemistry , azide , in vivo , substrate (aquarium) , in vitro , combinatorial chemistry , myristoylation , fusion protein , recombinant dna , enzyme , organic chemistry , membrane , biology , ecology , microbiology and biotechnology , gene
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.
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