N-Myristoyl transferase-mediated protein labelling in vivo |
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Authors: | Heal William P Wickramasinghe Sasala R Leatherbarrow Robin J Tate Edward W |
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Affiliation: | Department of Chemistry, South Kensington Campus, Imperial College, London, UK SW7 2AZ. |
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Abstract: | 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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