title: A fluorescent broad-spectrum proteasome inhibitor for labeling proteasomes in vitro and in vivo.
abstract: The proteasome is an essential evolutionary conserved protease involved in many regulatory systems. Here, we describe the synthesis and characterization of the activity-based, fluorescent, and cell-permeable inhibitor Bodipy TMR-Ahx(3)L(3)VS (MV151), which specifically targets all active subunits of the proteasome and immunoproteasome in living cells, allowing for rapid and sensitive in-gel detection. The inhibition profile of a panel of commonly used proteasome inhibitors could be readily determined by MV151 labeling. Administration of MV151 to mice allowed for in vivo labeling of proteasomes, which correlated with inhibition of proteasomal degradation in the affected tissues. This probe can be used for many applications ranging from clinical profiling of proteasome activity, to biochemical analysis of subunit specificity of inhibitors, and to cell biological analysis of the proteasome function and dynamics in living cells.
authors.affiliation: Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University, 2300 RA Leiden, The Netherlands.
authors.forename: Martijn
authors.initials: M
authors.lastname: Verdoes
authors.forename: Bogdan I
authors.initials: BI
authors.lastname: Florea
authors.forename: Victoria
authors.initials: V
authors.lastname: Menendez-Benito
authors.forename: Christa J
authors.initials: CJ
authors.lastname: Maynard
authors.forename: Martin D
authors.initials: MD
authors.lastname: Witte
authors.forename: Wouter A
authors.initials: WA
authors.lastname: van der Linden
authors.forename: Adrianus M C H
authors.initials: AM
authors.lastname: van den Nieuwendijk
authors.forename: Tanja
authors.initials: T
authors.lastname: Hofmann
authors.forename: Celia R
authors.initials: CR
authors.lastname: Berkers
authors.forename: Fijs W B
authors.initials: FW
authors.lastname: van Leeuwen
authors.forename: Tom A
authors.initials: TA
authors.lastname: Groothuis
authors.forename: Michiel A
authors.initials: MA
authors.lastname: Leeuwenburgh
authors.forename: Huib
authors.initials: H
authors.lastname: Ovaa
authors.forename: Jacques J
authors.initials: JJ
authors.lastname: Neefjes
authors.forename: Dmitri V
authors.initials: DV
authors.lastname: Filippov
authors.forename: Gijs A
authors.initials: GA
authors.lastname: van der Marel
authors.forename: Nico P
authors.initials: NP
authors.lastname: Dantuma
authors.forename: Herman S
authors.initials: HS
authors.lastname: Overkleeft
mesh_terms: D000818:Animals; D001896:Boron Compounds / Q000138:chemical synthesis / Q000494:pharmacology*; D045744:Cell Line, Tumor; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009842:Oligopeptides / Q000138:chemical synthesis / Q000494:pharmacology*; D011480:Protease Inhibitors / Q000138:chemical synthesis / Q000494:pharmacology*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D061988:Proteasome Inhibitors
chemical_list: C517407:Bodipy TMR-Ahx(3)L(3)VS; D001896:Boron Compounds; D005456:Fluorescent Dyes; D009842:Oligopeptides; D011480:Protease Inhibitors; D061988:Proteasome Inhibitors; D046988:Proteasome Endopeptidase Complex