Traceless Cleavage of Protein–Biotin Conjugates under Biologically Compatible Conditions. (19th July 2017)

• Record Type: Journal Article
• Authors: Cowell, Joseph; Buck, Matthew; Essa, Ali H.; Clarke, Rebecca; Vollmer, Waldemar; Vollmer, Daniela; Hilkens, Catharien M.; Isaacs, John D.; Hall, Michael J.; Gray, Joe
• Is Part Of: Chembiochem. Volume 18:Number 17(2017)
• Page Start: 1688
• Page End: 1691
• Publication Date: 2017-07-19
• Subjects: affinity purification -- protein modifications -- proteomics -- reversible biotinylation -- traceless cleavage; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.201700214
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: Biotinylation of amines is widely used to conjugate biomolecules, but either the resulting label is non‐removable or its removal leaves a tag on the molecule of interest, thus 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, room temperature) that regenerates the unmodified amine. The capture and release of biotinylated proteins and peptides from neutravidin, fluorescent labelling through 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. Abstract : Come undone : We demonstrate for the first time that it is possible to tracelessly regenerate biotin‐tagged biological amines (peptides and proteins) under extremely mild and biologically compatible conditions. These simple reagents offer new possibilities in the field of chemical biology, particularly in biorthogonal chemistry and proteomics.