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Na +/Ca2+ exchangers: relationship between structure and function

Grant number: 17/05614-8
Support type:Scholarships in Brazil - Doctorate (Direct)
Effective date (Start): August 01, 2017
Effective date (End): June 30, 2020
Field of knowledge:Biological Sciences - Biochemistry - Chemistry of Macromolecules
Principal researcher:Roberto Kopke Salinas
Grantee:Phelipe Augusto Mariano Vitale
Home Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil


Membrane proteins correspond to about 30% of the genes of eukaryotic and prokaryotic organisms, and approximately 60% of the targets for the development of new drugs. Nevertheless, the membrane proteins correspond to about 3% of the structures deposited in the PDB. Na+/Ca2+ (NCX) exchanger are membrane proteins absolutely essential for the maintenance of Ca2+ homeostasis in different cell types. They contain 10 transmembrane helices and a large cytoplasmic loop. The transmembrane domain is responsible for the transport of Na + and Ca2 + through the lipid bilayer, while the intracellular loop is responsible for the allosteric regulation of the exchanger: the Ca2+ binding in two sensor domains (CBD1 and CBD2) activates the NCX. This multifaceted character makes the structural characterization of NCX even more difficult. In fact, despite the recognized physiological importance, little is known about the three-dimensional structure of NCX, and the mechanism of allosteric regulation remains poorly understood. In this project we propose to combine functional and structural studies in order to obtain mechanistic information on the transport and regulation of the NCX exchangers. For this purpose we will carry out measurements of Ca2+ transport in eukaryotic cells involving wild and mutant NCX exchanger. We will produce prokaryotic and/or eukaryotic exchanger samples with the objective of carrying out structural studies by NMR in solution. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
DE SOUZA DEGENHARDT, MAXIMILIA F.; VITALE, PHELIPE A. M.; ABIKO, LAYARA A.; ZACHARIAS, MARTIN; SATTLER, MICHAEL; OLIVEIRA, CRISTIANO L. P.; SALINAS, ROBERTO K. Molecular insights on CALX-CBD12 interdomain dynamics from MD simulations, RDCs, and SAXS. BIOPHYSICAL JOURNAL, v. 120, n. 17, p. 3664-3675, SEP 7 2021. Web of Science Citations: 0.
V.V. DE SOUZA; P.A.M. VITALE; F.H. FLORENZANO; R.K. SALINAS; I.M. CUCCOVIA. A novel method for DNA delivery into bacteria using cationic copolymers. Brazilian Journal of Medical and Biological Research, v. 54, n. 5, p. -, 2021. Web of Science Citations: 0.
CARDOSO, MARCUS V. C.; RIVERA, JOSE D.; VITALE, PHELIPE A. M.; DEGENHARDT, MAXIMILIA F. S.; ABIKO, LAYARA A.; OLIVEIRA, CRISTIANO L. P.; SALINAS, ROBERTO K. CALX-CBD1 Ca2+-Binding Cooperativity Studied by NMR Spectroscopy and ITC with Bayesian Statistics. BIOPHYSICAL JOURNAL, v. 119, n. 2, p. 337-348, JUL 21 2020. Web of Science Citations: 0.

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