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Lookup NU author(s): Joseph Cowell, Matthew Buck, Rebecca Clarke, Professor Waldemar Vollmer, Dr Daniela Vollmer, Professor Catharien Hilkens, Professor John IsaacsORCiD, Dr Michael HallORCiD, Dr Joseph Gray
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
Biotinylation of amines is widely used to conjugate biomolecules, but the resulting label is either non-removable or removal leaves a tag on the molecule of interest, affecting downstream processes. We present here a set of reagents (RevAmines) that allow traceless, reversible biotinylation under biologically-compatible, mild conditions. Release following avidin-based capture is achieved through the cleavage of a (2-(alkylsulfonyl)ethyl) carbamate linker under mild conditions (200 mM ammonium bicarbonate, pH 8, 16 - 24 h, r.t.) regenerating the unmodified amine. The capture and release of biotinylated proteins and peptides from neutravidin, fluorescent labelling via reversible biotinylation at the cell surface and the selective enrichment of proteins from bacterial periplasm are demonstrated. The tags are easily prepared, stable and offer the potential for future application in proteomics, activity-based protein profiling, affinity chromatography and bio-molecule tagging and purification.
Author(s): Cowell J, Buck M, Essa AH, Clarke R, Vollmer W, Vollmer D, Hilkens CM, Isaacs JD, Hall MJ, Gray J
Publication type: Article
Publication status: Published
Journal: ChemBioChem
Year: 2017
Volume: 18
Issue: 17
Pages: 1688-1691
Print publication date: 05/09/2017
Online publication date: 05/06/2017
Acceptance date: 02/06/2017
Date deposited: 05/06/2017
ISSN (print): 1439-4227
ISSN (electronic): 1439-7633
Publisher: Wiley - V C H Verlag GmbH & Co. KGaA
URL: https://doi.org/10.1002/cbic.201700214
DOI: 10.1002/cbic.201700214
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