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

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

Abstract

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)

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)

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. (09/53989-4, 18/21450-8, 17/05614-8, 16/17375-5, 16/07490-1, 17/06394-1)

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, 2021-04-02. (17/05614-8, 16/07490-1, 13/08166-5)

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. (18/16092-5, 16/24531-3, 19/19968-1, 17/05614-8, 16/07490-1)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

VITALE, Phelipe Augusto Mariano. Na+/Ca2+ exchangers: Relationship between structure and function. 2022. Doctoral Thesis - Universidade de São Paulo (USP). Conjunto das Químicas (IQ e FCF) (CQ/DBDCQ) São Paulo.