A fluorescent broad-spectrum proteasome inhibitor for labeling proteasomes in vitro and in vivo.
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.
Verdoes
Martijn
M
Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University, 2300 RA Leiden, The Netherlands.
Florea
Bogdan I
BI
Menendez-Benito
Victoria
V
Maynard
Christa J
CJ
Witte
Martin D
MD
van der Linden
Wouter A
WA
van den Nieuwendijk
Adrianus M C H
AM
Hofmann
Tanja
T
Berkers
Celia R
CR
van Leeuwen
Fijs W B
FW
Groothuis
Tom A
TA
Leeuwenburgh
Michiel A
MA
Ovaa
Huib
H
Neefjes
Jacques J
JJ
Filippov
Dmitri V
DV
van der Marel
Gijs A
GA
Dantuma
Nico P
NP
Overkleeft
Herman S
HS
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
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