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Using isothermal titration calorimetry for the determination of thermodynamic properties of protein-ligand and protein-protein interactions

Grant number: 11/23110-0
Support type:Regular Research Grants
Duration: August 01, 2012 - July 31, 2014
Field of knowledge:Biological Sciences - Biophysics - Molecular Biophysics
Principal Investigator:Julio Cesar Borges
Grantee:Julio Cesar Borges
Home Institution: Instituto de Química de São Carlos (IQSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil

Abstract

Protein-protein and protein-ligand interactions are crucial events for the regulation of proteins and protein activity, for instance, it is responsible for enzyme activation or inhibition, by conformational changes in proteins for cell signaling, transport and storage of substances, etc. Few experimental techniques allow studying these types of interactions in solution, and provide a wide range of thermodynamic data such as isothermal titration calorimetry (ITC). This method uses heat, a universal probe, to yield information such as stoichiometry of the interaction, the interaction enthalpy, affinity constant (and change in free energy) and interaction entropy. Besides this information, ITC experiments at different temperatures allow to determine the change of heat capacity of interaction. Altogether, these data provide a "thermodynamic signature" of interaction that can assist in understanding the mechanism of interaction involved in the molecular level, especially with the aid of structural information obtained by other techniques. The molecular understanding of protein-ligand or protein-protein interactions may assist in understanding the mechanism of protein regulation and may also contribute to the rational drug design. The ITC also allows to monitor the flow of an enzymatic reaction producing information as a constant Michaelis-Menten, constant of catalysis and inhibition constant when performed in the presence of inhibitors. Thus, the objective of this project is to apply the ITC to obtain thermodynamic data for protein-ligand or protein-protein interactions to correlate this information with structural data, at low and high resolution, of several proteins in the study group BMB/IQSC/USP. The group BMB cover research in four main lines involving molecular chaperones Hsp90 and Hsp70 families (and their co-chaperones) from human and protozoa, also a trypanocidal-metabolizing enzyme. The ITC technique can considerably help the characterization of these proteins under study in the group BMB. (AU)

Scientific publications (17)
(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)
KIRALY, VANESSA T. R.; DORES-SILVA, PAULO R.; SERRAO, VITOR H. B.; CAUVI, DAVID M.; DE MAIO, ANTONIO; BORGES, JULIO C. Thermal aggregates of human mortalin and Hsp70-1A behave as supramolecular assemblies. International Journal of Biological Macromolecules, v. 146, p. 320-331, MAR 1 2020. Web of Science Citations: 0.
BATISTA, FERNANDA A. H.; RAMOS, JR., SERGIO L.; TASSONE, GIUSY; LEITAO, ANDREI; MONTANARI, CARLOS A.; BOTTA, MAURIZIO; MORI, MATTIA; BORGES, JULIO C. Discovery of small molecule inhibitors of Leishmania braziliensis Hsp90 chaperone. Journal of Enzyme Inhibition and Medicinal Chemistry, v. 35, n. 1, p. 639-649, JAN 1 2020. Web of Science Citations: 0.
SILVA, NOELI S. M.; BERTOLINO-REIS, DAYANE E.; DORES-SILVA, PAULO R.; ANNETA, FATIMA B.; SERAPHIM, THIAGO V.; BARBOSA, LEANDRO R. S.; BORGES, JULIO C. Structural studies of the Hsp70/Hsp90 organizing protein of Plasmodium falciparum and its modulation of Hsp70 and Hsp90 ATPase activities. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1868, n. 1 JAN 2020. Web of Science Citations: 0.
ALVES BARBOSA, EVERTON DE ALMEIDA; SERAPHIM, THIAGO VARGAS; GANDIN, CESAR AUGUSTO; TEIXEIRA, LEILANE FERREIRA; GONCALVES DA SILVA, RONNI ANDERSON; RIGHETTO, GERMANNA L.; GONCALVES, KALIANDRA DE ALMEIDA; VASCONCELLOS, RAPHAEL DE SOUZA; ALMEIDA, MARCIA ROGERIA; SILVA JUNIOR, ABELARDO; RANGEL FIETTO, JULIANA LOPES; KOBARG, JORG; GILEADI, CARINA; MASSIRER, KATLIN B.; BORGES, JULIO CESAR; NETO, MARIO DE OLIVEIRA; BRESSAN, GUSTAVO COSTA. Insights into the full-length SRPK2 structure and its hydrodynamic behavior. International Journal of Biological Macromolecules, v. 137, p. 205-214, SEP 15 2019. Web of Science Citations: 0.
MINARI, KARINE; DE AZEVEDO, ERIKA CHANG; RODRIGUES KIRALY, VANESSA THOMAZ; HELENO BATISTA, FERNANDA APARECIDA; DE MORAES, FABIO ROGERIO; DE MELO, FERNANDO ALVES; NASCIMENTO, ALESSANDRO SILVA; GAVA, LISANDRA MARQUES; INACIO RAMOS, CARLOS HENRIQUE; BORGES, JULIO CESAR. Thermodynamic analysis of interactions of the Hsp90 with adenosine nucleotides: A comparative perspective. International Journal of Biological Macromolecules, v. 130, p. 125-138, JUN 1 2019. Web of Science Citations: 1.
WALBERT VELOSO-SILVA, LAUDIMIR LEONARDO; DORES-SILVA, PAULO ROBERTO; BERTOLINO-REIS, DAYANE ELIARA; MORENO-OLIVEIRA, LOUIS FELLIPE; LIBARDI, SILVIA HELENA; BORGES, JULIO CESAR. Structural studies of Old Yellow Enzyme of Leishmania braziliensis in solution. Archives of Biochemistry and Biophysics, v. 661, p. 87-96, JAN 2019. Web of Science Citations: 1.
BATISTA, FERNANDA A. H.; DORES-SILVA, PAULO R.; BORGES, JULIO C. Molecular Chaperones Involved in Protein Recovery from Aggregates are Present in Protozoa Causative of Malaria and Leishmaniasis. CURRENT PROTEOMICS, v. 16, n. 1, p. 12-21, 2019. Web of Science Citations: 0.
SILVA, NOELI S. M.; SERAPHIM, THIAGO V.; MINARI, KARINE; BARBOSA, LEANDRO R. S.; BORGES, JULIO C. Comparative studies of the low-resolution structure of two p23 co-chaperones for Hsp90 identified in Plasmodium falciparum genome. International Journal of Biological Macromolecules, v. 108, p. 193-204, MAR 2018. Web of Science Citations: 3.
SERAPHIM, THIAGO V.; SILVA, KELLY P.; DORES-SILVA, PAULO R.; BARBOSA, LEANDRO R. S.; BORGES, JULIO C. Insights on the structural dynamics of Leishmania braziliensis Hsp90 molecular chaperone by small angle X-ray scattering. International Journal of Biological Macromolecules, v. 97, p. 503-512, APR 2017. Web of Science Citations: 6.
DORES-SILVA, PAULO R.; NISHIMURA, LETICIA S.; KIRALY, VANESSA T. R.; BORGES, JULIO C. Structural and functional studies of the Leishmania braziliensis mitochondrial Hsp70: Similarities and dissimilarities to human orthologues. Archives of Biochemistry and Biophysics, v. 613, p. 43-52, JAN 1 2017. Web of Science Citations: 2.
BATISTA, FERNANDA A. H.; SERAPHIM, THIAGO V.; SANTOS, CLELTON A.; GONZAGA, MARISVANDA R.; BARBOSA, LEANDRO R. S.; RAMOS, CARLOS H. I.; BORGES, JULIO C. Low sequence identity but high structural and functional conservation: The case of Hsp70/Hsp90 organizing protein (Hop/Sti1) of Leishmania braziliensis. Archives of Biochemistry and Biophysics, v. 600, p. 12-22, JUN 15 2016. Web of Science Citations: 3.
DORES-SILVA, P. R.; BELOTI, L. L.; MINARI, K.; SILVA, S. M. O.; BARBOSA, L. R. S.; BORGES, J. C. Structural and functional studies of Hsp70-escort protein-Hep1-of Leishmania braziliensis. International Journal of Biological Macromolecules, v. 79, p. 903-912, AUG 2015. Web of Science Citations: 4.
DORES-SILVA, PAULO R.; BARBOSA, LEANDRO R. S.; RAMOS, CARLOS H. I.; BORGES, JULIO C. Human Mitochondrial Hsp70 (Mortalin): Shedding Light on ATPase Activity, Interaction with Adenosine Nucleotides, Solution Structure and Domain Organization. PLoS One, v. 10, n. 1 JAN 23 2015. Web of Science Citations: 17.
SERAPHIM, THIAGO V.; GAVA, LISANDRA M.; MOKRY, DAVID Z.; CAGLIARI, THIAGO C.; BARBOSA, LEANDRO R. S.; RAMOS, CARLOS H. I.; BORGES, JULIO C. The C-terminal region of the human p23 chaperone modulates its structure and function. Archives of Biochemistry and Biophysics, v. 565, p. 57-67, JAN 1 2015. Web of Science Citations: 7.
BATISTA, FERNANDA A. H.; ALMEIDA, GLESSLER S.; SERAPHIM, THIAGO V.; SILVA, KELLY P.; MURTA, SILVANE M. F.; BARBOSA, LEANDRO R. S.; BORGES, J. ULIO C. Identification of two p23 co-chaperone isoforms in Leishmania braziliensis exhibiting similar structures and Hsp90 interaction properties despite divergent stabilities. FEBS Journal, v. 282, n. 2, p. 388-406, JAN 2015. Web of Science Citations: 11.
MURAKAMI, MARIO T.; RODRIGUES, NATHALIA C.; GAVA, LISANDRA M.; HONORATO, RODRIGO V.; CANDURI, FERNANDA; BARBOSA, LEANDRO R. S.; OLIVA, GLAUCIUS; BORGES, JULIO C. Structural studies of the Trypanosoma cruzi Old Yellow Enzyme: Insights into enzyme dynamics and specificity. Biophysical Chemistry, v. 184, p. 44-53, DEC 31 2013. Web of Science Citations: 4.
SERAPHIM, THIAGO V.; ALVES, MARINA M.; SILVA, INDJARA M.; GOMES, FRANCISCO E. R.; SILVA, KELLY P.; MURTA, SILVANE M. F.; BARBOSA, LEANDRO R. S.; BORGES, JULIO C. Low Resolution Structural Studies Indicate that the Activator of Hsp90 ATPase 1 (Aha1) of Leishmania braziliensis Has an Elongated Shape Which Allows Its Interaction with Both N- and M-Domains of Hsp90. PLoS One, v. 8, n. 6 JUN 24 2013. Web of Science Citations: 13.

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