title: First BRET-based screening assay performed in budding yeast leads to the discovery of CDK5/p25 interaction inhibitors.
abstract: The protein kinase CDK5 (cyclin-dependent kinase 5) is activated through its association with a cyclin-like protein p35 or p39. In pathological conditions (such as Alzheimer's disease and various other neuropathies), truncation of p35 leads to the appearance of the p25 protein. The interaction of p25 with CDK5 up-regulates the kinase activity and modifies the substrate specificity. ATP-mimetic inhibitors of CDK5 have already been developed. However, the lack of selectivity of such inhibitors is often a matter of concern. An alternative approach can be used to identify highly specific inhibitors that disrupt protein interactions involving protein kinases. We have developed a bioluminescence resonance energy transfer (BRET)-based screening assay in yeast to discover protein-protein interaction inhibitors (P2I2). Here, we present the first use of BRET in yeast for the screening of small molecule libraries. This screening campaign led to the discovery of one molecule that prevents the interaction between CDK5 and p25, thus inhibiting the protein kinase activity. This molecule may give rise to high-specificity drug candidates.
authors.affiliation: CNRS USR 3151, Protein Phosphorylation and Disease Laboratory, Protein-Protein Interaction Inhibition P2I2 Group, Station Biologique, Roscoff, Bretagne, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Corbel
authors.forename: Qian
authors.initials: Q
authors.lastname: Wang
authors.forename: Hadjira
authors.initials: H
authors.lastname: Bousserouel
authors.forename: Amel
authors.initials: A
authors.lastname: Hamdi
authors.forename: Bing
authors.initials: B
authors.lastname: Zhang
authors.forename: Olivier
authors.initials: O
authors.lastname: Lozach
authors.forename: Yoan
authors.initials: Y
authors.lastname: Ferandin
authors.forename: Vincent B C
authors.initials: VB
authors.lastname: Tan
authors.forename: Françoise
authors.initials: F
authors.lastname: Guéritte
authors.forename: Pierre
authors.initials: P
authors.lastname: Colas
authors.forename: Cyril
authors.initials: C
authors.lastname: Couturier
authors.forename: Stéphane
authors.initials: S
authors.lastname: Bach
mesh_terms: D000732:Androstanols / Q000494:pharmacology; D051360:Cyclin-Dependent Kinase 5 / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D055808:Drug Discovery / Q000379:methods*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000737:chemistry / Q000378:metabolism; D056004:Molecular Dynamics Simulation; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D054852:Small Molecule Libraries
chemical_list: D000732:Androstanols; D008164:Luminescent Proteins; D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins; D054852:Small Molecule Libraries; C089495:neuronal Cdk5 activator (p25-p35); D051360:Cyclin-Dependent Kinase 5