title: Two-step bioorthogonal activity-based proteasome profiling using copper-free click reagents: a comparative study.
abstract: The development and application of bioorthogonal two-step labeling techniques receives much attention. Employing bifunctional proteasome probe 2 the efficiency of two-step labeling of recently published biotinylated cyclooctynes 3-5 is compared to Staudinger-Bertozzi ligation in cell extracts and living cells. While cyclooctynes 3-5 react faster and at a much lower concentration then the Staudinger-Bertozzi benchmark, background labeling is considerable with these reagents.
authors.affiliation: Leiden Institute of Chemistry and Netherlands Proteomics Centre, PO Box 9502, 2300 RA, Leiden, The Netherlands.
authors.forename: Wouter A
authors.initials: WA
authors.lastname: van der Linden
authors.forename: Nan
authors.initials: N
authors.lastname: Li
authors.forename: Sascha
authors.initials: S
authors.lastname: Hoogendoorn
authors.forename: Mark
authors.initials: M
authors.lastname: Ruben
authors.forename: Martijn
authors.initials: M
authors.lastname: Verdoes
authors.forename: Jun
authors.initials: J
authors.lastname: Guo
authors.forename: Geert-Jan
authors.initials: GJ
authors.lastname: Boons
authors.forename: Gijsbert A
authors.initials: GA
authors.lastname: van der Marel
authors.forename: Bogdan I
authors.initials: BI
authors.lastname: Florea
authors.forename: Herman S
authors.initials: HS
authors.lastname: Overkleeft
mesh_terms: D001710:Biotin / Q000031:analogs & derivatives* / Q000737:chemistry; D002460:Cell Line; D057930:Click Chemistry; D003300:Copper / Q000737:chemistry; D034242:Cyclooctanes / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry / Q000378:metabolism*
chemical_list: D034242:Cyclooctanes; D005456:Fluorescent Dyes; D001710:Biotin; D003300:Copper; D046988:Proteasome Endopeptidase Complex