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Function Studies of Grb2 through Nuclear Magnetic Resonance and Fluorescence: correlating dynamics and structure

Grant number:19/24974-0
Support Opportunities:Regular Research Grants
Start date: May 01, 2020
End date: April 30, 2022
Field of knowledge:Physical Sciences and Mathematics - Physics
Principal Investigator:Fernando Alves de Melo
Grantee:Fernando Alves de Melo
Host Institution: Instituto de Biociências, Letras e Ciências Exatas (IBILCE). Universidade Estadual Paulista (UNESP). Campus de São José do Rio Preto. São José do Rio Preto , SP, Brazil
City of the host institution:São José do Rio Preto
Associated researchers:Fátima Pereira de Souza ; Ícaro Putinhon Caruso ; Marcelo Andrés Fossey

Abstract

Cell metabolism is mediated by signaling pathways that are up regulated through enzymatic amino acids side chain phosphorylation dependent. So Protein Tyrosine Kinases (PTKs) play pivotal role on gene expression, cellular growth, division and differentiation, among others. Once phosphorylated, PTKs recruits protein partners such as Grb2, Shc, Ras, etc. to build up early signaling complexes that will activate specific signaling pathways inside cell. In many cases kinases and phosphatases go through these processes together. FGFRs are key PTKs in many signaling process where the aberrant activity causes several human diseases, including cancer. The adaptor protein Grb2 (Growth-factor receptor bound protein 2) is an important regulator of FGFR2 and Shp2 by preventing kinase and phosphatase activity respectively before extra-cellular stimuli. Phosphorylation of Grb2 by FGFR2 abrogates its binding to the receptor, resulting in up-regulation of both FGFR2's kinase and Shp2's phosphatase activity. On the other hand, dephosphorylation of Grb2 by Shp2 rescues the FGFR2-Grb2 complex. Other study reports that Grb2 is a regulator of MAPK pathway activity. This study states that only monomeric Grb2 is capable of interacting to SOS that will result in an upregulation of MAPK signaling while the dimeric form of Grb2 is inhibitory to this process. Then, Grb2 is a global regulator of these mutually dependent reactions, which turn this protein in an important biological target to be studied and tested within anti-tumor molecules. Grb2's versatility on executing biological functions other than adaptor protein is probably related to its flexibility which is inherent to multi-domain and intrinsically disordered proteins so we intend to use NMR-based techniques to characterize Grb2's molecular dynamics, conformational changes induced by phosphorylation and/or binding and access intermediated folding states that could be related to cell proliferation. (AU)

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Scientific publications (5)
(The scientific publications listed on this page originate from the Web of Science or SciELO databases. Their authors have cited FAPESP grant or fellowship project numbers awarded to Principal Investigators or Fellowship Recipients, whether or not they are among the authors. This information is collected automatically and retrieved directly from those bibliometric databases.)
DIAS, RAPHAEL V. R.; PEDRO, RENAN P.; SANCHES, MURILO N.; MOREIRA, GIOVANA C.; LEITE, VITOR B. P.; CARUSO, ICARO P.; DE MELO, FERNANDO A.; DE OLIVEIRA, LEANDRO C.. Unveiling Metastable Ensembles of GRB2 and the Relevance of Interdomain Communication during Folding. JOURNAL OF CHEMICAL INFORMATION AND MODELING, v. 63, n. 20, p. 10-pg., . (19/24974-0, 14/17630-0, 21/15028-4, 19/22540-3, 23/02219-1, 16/08753-6)
TEDESCO, JESSICA A.; DIAS, RAPHAEL V. R.; CASTELUCI, GIOVANA; PEDRO, RENAN P.; DE OLIVEIRA, LEANDRO C.; CARUSO, ICARO P.; MELO, FERNANDO A.. The influence of pH on the structure and stability of the Grb2 dimer reveals changes in the inter-domain and molecular interaction: Could it be a modulation mechanism?. Biophysical Chemistry, v. 295, p. 11-pg., . (14/17630-0, 19/24974-0)
CASTELUCI, G.; DIAS, R. V. R.; MARTINS, I. B. S.; FERNANDES, R. A.; TEDESCO, J. A.; CARUSO, I. P.; DE ARAUJO, A. S.; ITRI, R.; MELO, F. A.. Grb2 Y160F mutant mimics the wild-type monomeric state dynamics and the monomer-dimer equilibrium. International Journal of Biological Macromolecules, v. 279, p. 9-pg., . (22/00347-0, 16/13884-2, 23/09642-7, 19/24974-0, 23/01632-2, 23/01744-5)
PEDRO, RENAN P.; DIAS, RAPHAEL V. R.; MARTINS, INGRID B. S.; NOGUEIRA SANCHES, MURILO; PILOTO, JOAO V.; CARUSO, ICARO P.; LEITE, VITOR B. P.; DE MELO, FERNANDO A.. Conformational Flexibility of GRB2 as a Key Factor in the Stability and Regulation of Its Interaction with SOS1. ACS OMEGA, v. 10, n. 27, p. 12-pg., . (21/15028-4, 14/17630-0, 23/01744-5, 22/07231-7, 23/09642-7, 23/01632-2, 19/24974-0, 22/13050-5, 23/02219-1)
SANCHES, KAROLINE; DIAS, RAPHAEL V. R.; DA SILVA, PAULO H.; CARUSO, ICARO P.; FOSSEY, MARCELO A.; DE SOUZA, FATIMA P.; DE OLIVEIRA, LEANDRO C.; MELO, FERNANDO A.. Thermodynamic profile and molecular modeling of the interaction between Grb2 dimer and flavonoids Rutin and Morin. Journal of Molecular Structure, v. 1234, p. 10-pg., . (16/08753-6, 14/17630-0, 19/24974-0)