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Traceless-Cleavage of Protein-Biotin Conjugates Under Biologically-Compatible Conditions

Lookup NU author(s): Joseph Cowell, Matthew Buck, Rebecca Clarke, Professor Waldemar Vollmer, Dr Daniela Vollmer, Dr Catharien Hilkens, Professor John Isaacs, Dr Michael Hall, Dr Joseph Gray

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

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.


Publication metadata

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

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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