Solution nuclear magnetic resonance spectroscopy applied to study protein dynamics

• Grant number: 16/17375-5
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: November 01, 2016
• End date: April 22, 2019
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Roberto Kopke Salinas
• Grantee: Marcus Vinícius Cangussu Cardoso
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Atomic-level understanding of basic biological phenomena, such as allostery, cooperativity and molecular recognition, depends on the availability of high-resolution structural information on the protein molecules involved. However, the protein native state is best described as a conformational ensemble and, hence, if one wants to understand how proteins carry out their biological functions it is necessary to describe not only the structure, but also the protein dynamics. Nuclear Magnetic Resonance is a well-suited spectroscopy technique to obtain detailed residue-level information on the protein internal motions. In this proposal, we will apply protein NMR methods to measure 15N relaxation parameters to unravel the internal motions of biological systems of our interest. Special attention will be paid to motions at slow time scales (ms to ms), which are particularly biologically relevant. We will use CPMG and/or T1r based relaxation experiments. Relaxation dispersion experiments may give relevant kinetic and thermodynamic information on conformational equilibriums of interest. Additionally, we will implement NMR pulse sequences to measure the 15N relaxation rates using TROSY. The proposed experiments will be applied to domains of the Na+/Ca2+ exchanger, a membrane protein that is involved in the maintenance of intracellular Ca2+ homeostasis. The same experiments will be also applied to study protein subunits of the Type IV Secretion System (T4SS) of Xanthomonas citri, a phytopathogen that infects citrus plants. (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)