Characterization of interactions in the protein quality system for the understanding of its role in the proteostasis maintenance

• Grant number: 17/26058-6
• Support type: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): August 01, 2018
• Effective date (End): January 23, 2019
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal researcher: Carlos Henrique Inacio Ramos
• Grantee: Dênio Emanuel Pires Souto
• Home Institution: Instituto de Química (IQ). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

Proteins are important cellular components, which are involved in almost all the physiological functions. In most cases, it is required that the protein folds to its native state and/or do not form aggregates in order to perform its functions. In the cell, a system composed of chaperones, Hsps and proteasomes, and known as PQC (Protein Quality Control) maintains the protein homeostasis and seems to play a central role in important diseases, such as some types of cancer and neurodegenerative diseases (e.g. Parkinson and Alzheimer's). It is recognized that the components of the PQC system interact with several proteins for the proteostasis maintenance, besides interacting with each other to increase the efficiency of the system. However, little is still known about the characteristics of these interactions due to the substantial number of components and its diversity between organisms and also between different organelles. In this context, the main goal of this proposal is to combine two complementary techniques in the characterization of protein-protein interaction, Surface Plasmon Resonance (SPR) and Calorimetry (Titration and Scanning), to investigate the interactions between the PQC system components and between these and the targets. This strategy benefits from the expertise of the candidate and supervisor in this research area, which are also complementary. Initially, the interactions between the Hsp70 and Hsp90 chaperones with their co-chaperones and target proteins will be investigated. As proteins targeted by these chaperones, we chose proteins from viruses that cause numerous impacting diseases in Brazil, such as Zika, Chikungunya and yellow fever, since both Hsp70 and Hsp90 help in the folding these protein in the cellular environment. It is important to mention that the plasmonic is a well-established field of research that indicates SPR as one of the main tools to study biomolecular interactions. In this proposal, SPR-based biosensor strategies will be developed for detailed quantitative analysis, through kinetic and thermodynamic approaches, of the biochemical events resulted from the investigated interactions. It is expected that very useful information will be obtained for a better understanding of the biochemical mechanisms that may be associated with the function of PQC system, including the human Hsp70/Hsp90 system on the mentioned viral proteins. In general terms, the present proposal addresses a subject of enormous relevance for the country and for the science, with great potential to yield fascinating discoveries that lead to the generation of new strategies aimed at therapeutic intervention and, therefore, deserves to be investigated with high priority.

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)

GONCALVES, CONRADO DE C.; PINHEIRO, GLAUCIA M. S.; DAHLSTROM, KATHE M.; SOUTO, DENIO E. P.; KUBOTA, LAURO T.; BARBOSA, LEANDRO R. S.; RAMOS, I, CARLOS H. On the structure and function of Sorghum bicolor CHIP (carboxyl terminus of Hsc70-interacting protein): A link between chaperone and proteasome systems. Plant Science, v. 296, JUL 2020. Web of Science Citations: 0.

SOUTO, DENIO E. P.; VOLPE, JAQUELINE; GONCALVES, CONRADO DE C.; RAMOS, I, CARLOS H.; KUBOTA, LAURO T. A brief review on the strategy of developing SPR-based biosensors for application to the diagnosis of neglected tropical diseases. Talanta, v. 205, DEC 1 2019. Web of Science Citations: 1.

YAMIL CHAIN, CECILIA; PIRES SOUTO, DENIO EMANUEL; LAURA SBARAGLINI, MARIA; LABRIOLA, CARLOS A.; DAZA MILLONE, MARIA ANTONIETA; ALEJANDRO RAMIREZ, EDUARDO; SEBASTIAN CISNEROS, JOSE; LOPEZ-ALBIZU, CONSTANZA; SCOLLO, KARENINA; KUBOTA, LAURO T.; MARIANO RUIZ, ANDRES; ELENA VELA, MARIA. Trypanosoma cruzi Virulence Factors for the Diagnosis of Chagas' Disease. ACS INFECTIOUS DISEASES, v. 5, n. 11, p. 1813-1819, NOV 2019. Web of Science Citations: 0.