title: Probing the proteasome cavity in three steps: bio-orthogonal photo-reactive suicide substrates.
abstract: Tri-functional activity-based protein probes that encompass an electrophilic trap, a photo-reactive group and a bio-orthogonal ligation handle are described. With these, and in a three-step chemical proteomics approach, proteasomal catalytic sites are covalently and irreversibly modified, followed by photocrosslinking of these to flanking subunits and Staudinger-Bertozzi ligation for visualization and identification of the resulting conjugates.
authors.affiliation: Leiden Institute of Chemistry and Netherlands Proteomics Centre, Gorlaeus Laboratories, Einsteinweg 55, 2333 CC Leiden, The Netherlands.
authors.forename: Paul P
authors.initials: PP
authors.lastname: Geurink
authors.forename: Bogdan I
authors.initials: BI
authors.lastname: Florea
authors.forename: Gijs A
authors.initials: GA
authors.lastname: Van der Marel
authors.forename: Benedikt M
authors.initials: BM
authors.lastname: Kessler
authors.forename: Herman S
authors.initials: HS
authors.lastname: Overkleeft
mesh_terms: D000818:Animals; D020134:Catalytic Domain; D045744:Cell Line, Tumor; D051379:Mice; D015335:Molecular Probes / Q000737:chemistry; D055668:Photochemical Processes; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*
chemical_list: D015335:Molecular Probes; D046988:Proteasome Endopeptidase Complex