N-myristoyl transferase-mediated protein labelling in vivo

被引:109
作者
Heal, William P. [1 ]
Wickramasinghe, Sasala R. [1 ]
Leatherbarrow, Robin J. [1 ,2 ]
Tate, Edward W. [1 ,2 ]
机构
[1] Univ London Imperial Coll Sci Technol & Med, Dept Chem, London SW7 2AZ, England
[2] Univ London Imperial Coll Sci Technol & Med, Chem Biol Ctr, London SW7 2AZ, England
基金
英国医学研究理事会; 英国生物技术与生命科学研究理事会;
关键词
D O I
10.1039/b803258k
中图分类号
O62 [有机化学];
学科分类号
070303 [有机化学]; 081704 [应用化学];
摘要
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.
引用
收藏
页码:2308 / 2315
页数:8
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