Proteasome selectivity towards Michael acceptor containing oligopeptide-based inhibitors.
The synthesis and biological evaluation of ten Michael acceptors containing potential proteasome inhibitors are described. Cellular targets of azide containing inhibitors and were assessed in HEK293T and RAW264.7 cells by a two step labeling strategy, followed by biotin-pulldown, affinity purification, on-bead tryptic digestion and LC-MS(2) identification. This strategy appears to be an attractive alternative to gel-based competition assays.
van der Linden
Wouter A
WA
Leiden Institute of Chemistry and Netherlands Proteomics Centre, P.O. Box 9502, 2300 RA, Leiden, The Netherlands.
Geurink
Paul P
PP
Oskam
Chris
C
van der Marel
Gijsbert A
GA
Florea
Bogdan I
BI
Overkleeft
Herman S
HS
D000818:Animals; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D006801:Humans; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D061988:Proteasome Inhibitors
D004791:Enzyme Inhibitors; D009842:Oligopeptides; D061988:Proteasome Inhibitors; D046988:Proteasome Endopeptidase Complex