2 research outputs found
Noncovalent Fluorescent Probes of Human Immuno- and Constitutive Proteasomes
We report here the synthesis and
biological evaluation of fluorescent
probes functioning as inhibitors that noncovalently block human immuno-
and constitutive proteasomes. These cell-penetrating linear analogues
of the natural cyclopeptide TMC-95A were efficient on cells at the
nanomolar level and assessed by confocal microscopy and flow cytometry.
They may constitute an alternative to previously reported fluorescent
probes that all bind covalently to proteasomes
Dimerized Linear Mimics of a Natural Cyclopeptide (TMC-95A) Are Potent Noncovalent Inhibitors of the Eukaryotic 20S Proteasome
Noncovalent
proteasome inhibitors introduce an alternative mechanism
of inhibition to that of covalent inhibitors used in cancer therapy.
Starting from a noncovalent linear mimic of TMC-95A, a series of dimerized
inhibitors using polyaminohexanoic acid spacers has been designed
and optimized to target simultaneously two of the six active sites
of the eukaryotic 20S proteasome. The homodimerized compounds actively
inhibited chymotrypsin-like (<i>K</i><sub>i</sub> = 6ā11
nM) and trypsin-like activities, whereas postacid activity was poorly
modified. The noncovalent binding mode was ascertained by X-ray crystallography
of the inhibitors complexed with the yeast 20S proteasome. The inhibition
of proteasomal activities in human cells was evaluated. The use of
the multivalency inhibitor concept has produced highly efficient and
selective noncovalent compounds (no inhibition of calpain and cathepsin)
that have potential therapeutic advantages compared to covalent binders
such as bortezomib and carfilzomib