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Chem Sci. 2013 Jul 29;4(9):3455-3458. Epub 2013 Jun 18.

A novel approach to the site-selective dual labelling of a protein via chemoselective cysteine modification.

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Department of Chemistry , University College London , 20 Gordon Street , London , WC1H OAJ , UK . Email: ; ; Tel: +44 (0)20 3108 5071.


Local protein microenvironment is used to control the outcome of reaction between cysteine residues and 2,5-dibromohexanediamide. The differential reactivity is exploited to introduce two orthogonal reactive handles onto the surface of a double cysteine mutant of superfolder green fluorescent protein in a regioselective manner. Subsequent elaboration with commonly used thiol and alkyne containing reagents affects site-selective protein dual labelling.

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