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Identification of Novel Classes of Protein Kinase Inhibitors Using Combinatorial Peptide Chemistry Based on Functional Genomics Knowledge
journal contribution
posted on 1999-02-24, 00:00 authored by Thomas J. Lukas, Salida Mirzoeva, Urszula Slomczynska, D. Martin WattersonA discovery approach based on an intramolecular inhibitory mechanism was applied to a
prototype calmodulin (CaM)-regulated protein kinase in order to demonstrate a proof-of-principle
for the development of selective inhibitors. The overall approach used functional genomics
analysis of myosin light chain kinase (MLCK) to identify short autoinhibitory sequences that
lack CaM recognition activity, followed by recursive combinatorial peptide library production
and comparative activity screens. Peptide 18 (Arg-Lys-Lys-Tyr-Lys-Tyr-Arg-Arg-Lys-NH2), one
of several selective inhibitors discovered, has an IC50 = 50 nM for MLCK, inhibits CaM kinase
II only at 4000-fold higher concentrations, and does not inhibit cyclic AMP-dependent protein
kinase. Analogues of peptide 18 containing conformationally constrained cis-4-aminocyclohexanecarboxylic acid retained affinity and selectivity. The inhibitors add to the armamentarium
available for the deconvolution of complex signal transduction pathways and their relationship
to homeostasis and disease, and the approach is potentially applicable to enzymes in which
the catalytic and regulatory domains are found within the same open reading frame of a cDNA.
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Novel ClassesCaM kinase IImyosin light chain kinasereading frame50 nMactivity screensgenomics analysisdiscovery approachautoinhibitory sequencesProtein Kinase Inhibitorsprototype calmodulincombinatorial peptide library productionCombinatorial Peptide ChemistryMLCKPeptide 18IC 50proteinpeptide 18lack CaM recognition activitysignal transduction pathwaysFunctional Genomics Knowledge
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