title: Protein enrichment by capture-release based on strain-promoted cycloaddition of azide with bicyclononyne (BCN).
abstract: An enrichment strategy was devised for azide derivatized macromolecules, based on strain-promoted alkyne-azide cycloaddition (SPAAC) and a cleavable linker. A ring-strained alkyne, bicyclo[6.1.0]non-4-yne (BCN), was covalently attached to agarose beads via a hydrazine-sensitive linker. Benchmark studies of the resulting 'azido-trap' beads were performed with a fluorogenic coumarin derivative, leading to efficient capture of the azidocoumarin with concomitant fluorescence staining of the beads via SPAAC. The versatility of the beads for specific protein enrichment was shown by an effective and highly specific capture-release strategy for enrichment of azido-containing Candida antarctica lipase B (CalB) from a mixture of proteins. This approach is suited for selective enrichment of (glyco)proteins after metabolic incorporation of azides for subsequent (glyco)proteomics studies.
authors.affiliation: Institute for Molecules and Materials, Radboud University Nijmegen, Heijendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.
authors.forename: Rinske P
authors.initials: RP
authors.lastname: Temming
authors.forename: Monique
authors.initials: M
authors.lastname: van Scherpenzeel
authors.forename: Esra
authors.initials: E
authors.lastname: te Brinke
authors.forename: Sanne
authors.initials: S
authors.lastname: Schoffelen
authors.forename: Jolein
authors.initials: J
authors.lastname: Gloerich
authors.forename: Dirk J
authors.initials: DJ
authors.lastname: Lefeber
authors.forename: Floris L
authors.initials: FL
authors.lastname: van Delft
mesh_terms: D000480:Alkynes / Q000737:chemistry; D001386:Azides / Q000737:chemistry*; D001643:Bridged Bicyclo Compounds / Q000737:chemistry*; D002175:Candida / Q000201:enzymology; D002384:Catalysis; D057930:Click Chemistry; D003500:Cyclization; D005456:Fluorescent Dyes / Q000737:chemistry; D005656:Fungal Proteins; D008049:Lipase / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000480:Alkynes; D001386:Azides; D001643:Bridged Bicyclo Compounds; D005456:Fluorescent Dyes; D005656:Fungal Proteins; C556617:bicyclo(6.1.0)nonyne; D008049:Lipase; C431349:lipase B, Candida antarctica