Interaction mechanism between the co-chaperone J-domain protein with their substrates and the chaperone Hsp70

• Grant number: 18/11948-9
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: November 01, 2018
• End date: December 31, 2021
• Field of knowledge: Physical Sciences and Mathematics - Chemistry - Organic Chemistry
• Principal Investigator: Carlos Henrique Inacio Ramos
• Grantee: Glaucia Melina Squizato Pinheiro de Castro
• Host Institution: Instituto de Química (IQ). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

Protein folding in the cell is usually aided by molecular chaperones, which the Hsp70 family (Hsp = heat shock protein) plays important roles, aiding in the folding of newly synthesized proteins, in the translocation, and others. Co-chaperones J-domain proteins (JDP) cooperate with Hsp70, binding to partially folded proteins (client) and delivering them to Hsp70. Although the functions of these proteins are determined, their three-dimensional structures and the interaction mechanism of JDP with the client and with Hsp70, for being transient, are not well established. Besides that, the absence of data is because the structural characterization of these proteins is hampered by the high molecular mass of these macromolecules, the presence of a large flexible region in the JDP and also because this interaction is transient. However, our research group has made important contributions about the structure and function of some JDP, characterizing the quaternary structure by low-angle scattering - SAXS (Ramos et al., 2008), and solving the three-dimensional NMR structure of the functional domain of JDP, which interacts with Hsp70 (Pinheiro et al., 2018). Also performed dynamics experiments, to understand the relationship between the domains, and which residues are involved in the interaction with the Hsp70 (manuscript in preparation). Since then we have been planning and executing experiments to solve the structure and dynamics of this co-chaperone in high resolution. The initial data obtained are very promising and suggest that we are close to accomplishing a feat in Brazil, determining the high resolution NMR structure of a dimeric protein, where each monomer has about 40 kDa and the understanding the interaction mechanism between this protein and the Hsp70 by cryo-electron microscopy. To reach this challenge, the collaboration between research groups at UNICAMP, UFRJ and IBS (Grenoble - France) is fundamental and such collaboration will be possible only through the candidate for the fellowships.

Scientific publications

(4)

(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)

PINHEIRO, GLAUCIA M. S.; AMORIM, GISELE C.; IQBAL, ANWAR; ALMEIDA, FABIO C. L.; RAMOSA, C. H. I.. Solution NMR investigation on the structure and function of the isolated J-domain from Sis1: Evidence of transient inter-domain interactions in the full-length protein. Archives of Biochemistry and Biophysics, v. 669, p. 71-79, JUL 15 2019. (17/26131-5, 12/50161-8, 17/01074-9, 18/11948-9)

AYINDE, KEHINDE S.; PINHEIRO, GLAUCIA M. S.; RAMOS, CARLOS H. I.. Binding of SARS-CoV-2 protein ORF9b to mitochondrial translocase TOM70 prevents its interaction with chaperone HSP90. Biochimie, v. 200, p. 8-pg., 2022-09-01. (17/26131-5, 18/11948-9)

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. (17/26158-0, 18/11948-9, 17/26058-6, 14/00076-0, 16/14228-1, 12/50161-8, 15/15822-1)

FRANCO, JULIANA C.; NOGUEIRA, MARIA L. C.; GANDELINI, GABRIELA M.; PINHEIRO, GLAUCIA M. S.; GONCALVES, CONRADO C.; BARBOSA, LEANDRO R. S.; YOUNG, JASON C.; RAMOS, CARLOS H. I.. Sorghum bicolor SbHSP110 has an elongated shape and is able of protecting against aggregation and replacing human HSPH1/HSP110 in refolding and disaggregation assays. Biopolymers, v. 114, n. 2, p. 12-pg., 2023-02-01. (16/02137-1, 12/50161-8, 16/14503-2, 16/04246-2, 16/03764-0, 18/11948-9, 17/26131-5)