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.
机构:Washington Univ, Sch Med, Dept Mol Biol & Pharmacol, St Louis, MO 63110 USA
Farazi, TA
;
Waksman, G
论文数: 0引用数: 0
h-index: 0
机构:Washington Univ, Sch Med, Dept Mol Biol & Pharmacol, St Louis, MO 63110 USA
Waksman, G
;
Gordon, JI
论文数: 0引用数: 0
h-index: 0
机构:
Washington Univ, Sch Med, Dept Mol Biol & Pharmacol, St Louis, MO 63110 USAWashington Univ, Sch Med, Dept Mol Biol & Pharmacol, St Louis, MO 63110 USA
机构:Washington Univ, Sch Med, Dept Mol Biol & Pharmacol, St Louis, MO 63110 USA
Farazi, TA
;
Waksman, G
论文数: 0引用数: 0
h-index: 0
机构:Washington Univ, Sch Med, Dept Mol Biol & Pharmacol, St Louis, MO 63110 USA
Waksman, G
;
Gordon, JI
论文数: 0引用数: 0
h-index: 0
机构:
Washington Univ, Sch Med, Dept Mol Biol & Pharmacol, St Louis, MO 63110 USAWashington Univ, Sch Med, Dept Mol Biol & Pharmacol, St Louis, MO 63110 USA