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Conformational antibodies for cPKCs and their applications

Grant number: 14/16037-3
Support type:Scholarships abroad - Research Internship - Doctorate
Effective date (Start): November 01, 2014
Effective date (End): August 31, 2015
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal researcher:Deborah Schechtman
Grantee:Darlene Aparecida Pena
Supervisor abroad: Lakshmi A. Devi
Home Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Research place: Icahn School of Medicine at Mount Sinai, United States  
Associated to the scholarship:11/10321-3 - Functional characterization of protein kinase c beta 1 in self-renewal, BP.DR


The Protein Kinase C (PKC) family of serine/ treonine quinases can be divided according to their activation requirements in classical PKCs (cPKCs) that include PKC alpha,beta I, beta II, and gamma, novel PKCs (nPKCs), including PKC delta, PKC epsilon, PKC eta and PKC theta, and atypical PKCs (aPKCs), PKC zeta and lambda/iota. During activation, PKC translocates to membranes and suffers several conformational changes. There are difficulties in specifically studying and detecting active PKCs, becoming a challenge to study their function. Specifically for cPKCs, phosphorylation does not correlate with activation but rather binding to lipids. Thus, during my PhD project (FAPESP# 2011/10321-3) we developed conformational antibodies against active forms of cPKC and have validated them in a cancer model, detecting binding partners of cPKCs in a metastatic cancer cell line. In the present study, we will collaborate with Dr. Lakshmi Devi an expert on conformational antibodies and drug addiction, working at the Center for Systems Biology at Mount Sinai, to further characterize our conformational antibodies and test them in an opiate addiction model. We propose to use these antibodies to identify PKC-associated regulatory synaptic networks during drug addiction. Several studies indicate a critical role for PKC, and specifically PKC gamma, in drug addiction. Identifying PKC gamma substrates and binding proteins of active PKC can help us elucidate the signaling pathways that are activated in chronic morphine treatment. (AU)

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