Identification of Protein Kinase C Activation as a Novel Mechanism for RGS2 Protein Upregulation through Phenotypic Screening of Natural Product Extracts

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dc.creatorRaveh, Avi
dc.creatorSchultz, Pamela J.
dc.creatorAschermann, Lauren 
dc.creatorCarpenter, Colleen
dc.creatorTamayo Castillo, Giselle 
dc.creatorCao, Shugeng
dc.creatorClardy, Jon
dc.creatorNeubig, Richard R.
dc.creatorSherman, David H.
dc.creatorSjögren, Benita
dc.date.accessioned2023-01-03T15:23:23Z
dc.date.available2023-01-03T15:23:23Z
dc.date.issued2014
dc.description.abstractBiochemical high-throughput screening is widely used in drug discovery, using a variety of small molecule libraries. However, broader screening strategies may be more beneficial to identify novel biologic mechanisms. In the current study we used a β-galactosidase complementation method to screen a selection of microbial-derived pre-fractionated natural product extracts for those that increase regulator of G protein signaling 2 (RGS2) protein levels. RGS2 is a member of a large family of proteins that all regulate signaling through G protein–coupled receptors (GPCRs) by accelerating GTPase activity on active Gα as well as through other mechanisms. RGS2−/− mice are hypertensive, show increased anxiety, and are prone to heart failure. RGS2 has a very short protein half-life due to rapid proteasomal degradation, and we propose that enhancement of RGS2 protein levels could be a beneficial therapeutic strategy. Bioassay-guided fractionation of one of the hit strains yielded a pure compound, Indolactam V, a known protein kinase C (PKC) activator, which selectively increased RGS2 protein levels in a time- and concentration-dependent manner. Similar results were obtained with phorbol 12-myristate 13-acetate as well as activation of the Gq-coupled muscarinic M3 receptor. The effect on RGS2 protein levels was blocked by the nonselective PKC inhibitor Gö6983 (3-[1-[3-(dimethylamino)propyl]-5-methoxy-1H-indol-3-yl]-4-(1H-indol-3-yl)-1H-pyrrole-2,5-dione), the PKCβ-selective inhibitor Ruboxastaurin, as well as small interfering RNA-mediated knockdown of PKCβ. Indolactam V-mediated increases in RGS2 protein levels also had functional effects on GPCR signaling. This study provides important proof-of-concept for our screening strategy and could define a negative feedback mechanism in Gq/Phospholipase C signaling through RGS2 protein upregulation.es_ES
dc.description.procedenceUCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de Químicaes_ES
dc.description.procedenceUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigaciones en Productos Naturales (CIPRONA)es_ES
dc.identifier.citationhttps://molpharm.aspetjournals.org/content/86/4/406.shortes_ES
dc.identifier.doi10.1124/mol.114.092403
dc.identifier.issn1521-0111
dc.identifier.urihttps://hdl.handle.net/10669/87982
dc.language.isoenges_ES
dc.rightsacceso embargado
dc.sourceMolecular Pharmacology, 86(4), p. 406-416es_ES
dc.subjectPROTEINSes_ES
dc.subjectBIOCHEMICALSes_ES
dc.subjectBIOCHEMICAL ANALYSISes_ES
dc.titleIdentification of Protein Kinase C Activation as a Novel Mechanism for RGS2 Protein Upregulation through Phenotypic Screening of Natural Product Extractses_ES
dc.typeartículo originales_ES

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