Studies of the mitochondrial HSP70 of human and protozoa: structural and functional approaches

• Grant number: 14/07206-6
• Support Opportunities: Regular Research Grants
• Start date: July 01, 2015
• End date: June 30, 2017
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Julio Cesar Borges
• Grantee: Julio Cesar Borges
• Host Institution: Instituto de Química de São Carlos (IQSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil

Abstract

Hsp70 is a major molecular chaperones and function as pivot receiving and distributing substrates from/to other molecular chaperones, both in the cytoplasm and in organelles. The mitochondrial Hsp70 (mtHsp70) is involved in the import of mitochondrial proteins produced in the cytoplasm and their subsequent folding, is essential for mitochondrial biogenesis. In mammals, the mtHsp70 is called mortalin due to its involvement in apoptosis and senescence. There is great interest in the mortalin study since it is overexpressed in tumor cells, which makes it as target for pharmacological inhibition, besides participating in the maintenance of various diseases caused by aging, such as Parkinson's and Alzheimer's. Although known for a long time, the production of recombinant human mortalin is limited due to a self-aggregation process which results in their recombinant production in aggregated form. Recently, a new mtHsp70 co-chaperone was described that operates assisting in the mtHsp70 correct folding. This mitochondrial protein is called Hep1 (Hsp70 escort protein 1) and allows the obtaining of recombinant mtHsp70 soluble and in the functional form. One goal of this project is the co-expression of human mortalin and Hep1 (hHep1) to enable the production of mortalin soluble as well as the structural and functional characterization of recombinant human mortalin. Objectives of this proposal are also the characterization of Leishmania braziliensis orthologue Hep1 (LbHep1) that has shown activity in maintaining human mortalin as well as mtHsp70 L. braziliensis (LbmtHsp70) in their soluble forms. From this study, it is expected (1) understanding the functional mechanism of the interaction with the Hep1 and mortalin and infer other possible functions for this protein. With getting the mortalin in the soluble form, is also expected (2) to obtain structural and functional information of this protein to enable compare it to cytoplasmic Hsp70. It is also expected (3) characterize the interaction of mortalin with its human nucleotide exchange factors (hGrpE) of mitochondrial origin, hGrpe # 1 and # 2. Regarding the protozoa systems, (4) the objective is to study in more detail the mechanism of interaction of LbHep1 with LbmtHsp70 , and ( 5 ) the operation of LbmtHsp70 comparing it to human mortalin. (AU)

Scientific publications

(20)

(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)

COTO, AMANDA L. S.; SERAPHIM, THIAGO V.; BATISTA, FERNANDA A. H.; DORES-SILVA, PAULO R.; BARRANCO, ANA BEATRIZ F.; TEIXEIRA, FELIPE R.; GAVA, LISANDRA M.; BORGES, JULIO C.. Structural and functional studies of the Leishmania braziliensis SGT co-chaperone indicate that it shares structural features with HIP and can interact with both Hsp90 and Hsp70 with similar affinities. International Journal of Biological Macromolecules, v. 118, n. A, p. 693-706, OCT 15 2018. (12/50161-8, 14/07206-6, 17/07335-9)

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. (11/23110-0, 12/50161-8, 14/07206-6)

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. (13/10712-8, 17/26131-5, 14/07206-6, 11/23110-0, 13/18009-4, 12/50161-8, 17/07335-9)

ROCHA, MARINA CAMPOS; MINARI, KARINE; FABRI, JOAO HENRIQUE TADINI MARILHANO; KERKAERT, JOSHUA D.; GAVA, LISANDRA MARQUES; DA CUNHA, ANDERSON FERREIRA; CRAMER, ROBERT A.; BORGES, JULIO CESAR; MALAVAZI, IRAN. Aspergillus fumigatusHsp90 interacts with the main components of the cell wall integrity pathway and cooperates in heat shock and cell wall stress adaptation. Cellular Microbiology, v. 23, n. 2, FEB 2021. (16/07870-9, 14/07206-6, 17/19694-3, 15/17541-0, 17/26131-5, 17/07335-9)

COTO, AMANDA L. S.; SERAPHIM, THIAGO V.; BATISTA, FERNANDA A. H.; DORES-SILVA, PAULO R.; BARRANCO, ANA BEATRIZ F.; TEIXEIRA, FELIPE R.; GAVA, LISANDRA M.; BORGES, JULIO C.. Structural and functional studies of the Leishmania braziliensis SGT co-chaperone indicate that it shares structural features with HIP and can interact with both Hsp90 and Hsp70 with similar affinities. International Journal of Biological Macromolecules, v. 118, p. 14-pg., 2018-10-15. (12/50161-8, 17/07335-9, 14/07206-6)

SILVA, NOELI S. M.; TORRICILLAS, MARCELA S.; MINARI, KARINE; BARBOSA, LEANDRO R. S.; SERAPHIM, V, THIAGO; BORGES, JULIO C.. Solution structure of Plasmodium falciparum Hsp90 indicates a high flexible dimer. Archives of Biochemistry and Biophysics, v. 690, SEP 15 2020. (14/07206-6, 17/26131-5, 11/23110-0, 17/07335-9, 12/50161-8)

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; et al. Insights into the full-length SRPK2 structure and its hydrodynamic behavior. International Journal of Biological Macromolecules, v. 137, p. 205-214, SEP 15 2019. (14/07206-6, 13/50724-5, 11/23110-0, 12/00195-3, 17/03489-1, 12/50161-8, 17/07335-9)

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. (17/26131-5, 14/07206-6, 11/23110-0, 12/50161-8, 17/07335-9, 14/16646-0)

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. (15/26722-8, 13/25646-0, 17/18173-0, 11/23110-0, 14/07206-6, 12/50161-8, 09/53989-4, 17/07335-9)

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. (11/23110-0, 14/07206-6, 17/07335-9)

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. (11/23110-0, 14/07206-6, 14/16646-0, 12/50161-8)

MELO SILVA, NOELI SOARES; DE CAMARGO RODRIGUES, LUIZ FERNANDO; DORES-SILVA, PAULO ROBERTO; MONTANARI, CARLOS ALBERTO; INACIO RAMOS, CARLOS HENRIQUE; SOUZA BARBOSA, LEANDRO RAMOS; BORGES, JULIO CESAR. Structural, thermodynamic and functional studies of human 71 kDa heat shock cognate protein (HSPA8/hHsc70). BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1869, n. 12, DEC 2021. (17/07335-9, 11/23110-0, 17/26131-5, 12/50161-8, 15/15822-1, 14/07206-6, 14/16646-0)

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. (17/26131-5, 14/07206-6, 11/23110-0, 12/50161-8, 17/07335-9)

DORES-SILVA, PAULO ROBERTO; RODRIGUES KIRALY, VANESSA THOMAZ; DE OLIVEIRA MORITZ, MILENE NOBREGA; BALASCO SERRAO, VITOR HUGO; SIQUEIRA DOS PASSOS, PATRICIA MARIA; SPAGNOL, VALENTINE; TEIXEIRA, FELIPE ROBERTI; GAVA, LISANDRA MARQUES; CAUVI, DAVID MARIO; INACIO RAMOS, CARLOS HENRIQUE; et al. New insights on human Hsp70-escort protein 1: Chaperone activity, interaction with liposomes, cellular localizations and HSPA's self-assemblies remodeling. International Journal of Biological Macromolecules, v. 182, p. 772-784, JUL 1 2021. (14/16646-0, 14/07206-6, 17/26131-5, 16/25798-3, 09/54216-9, 12/50161-8, 17/07879-9, 11/23110-0, 17/07335-9, 12/23730-1)

MOHAMMADI-OSTAD-KALAYEH, SONA; STAHL, FRANK; SCHEPER, THOMAS; KOCK, KLAUS; HERRMANN, CHRISTIAN; BATISTA, FERNANDA APARECIDA HELENO; BORGES, JULIO CESAR; SASSE, FLORENZ; EICHNER, SIMONE; ONGOUTA, JEKATERINA; et al. Heat Shock Proteins Revisited: Using a Mutasynthetically Generated Reblastatin Library to Compare the Inhibition of Human and Leishmania Hsp90s. CHEMBIOCHEM, v. 19, n. 6, p. 562-574, MAR 16 2018. (12/50161-8, 14/07206-6)

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. (14/07206-6, 13/25646-0, 11/23110-0, 12/50161-8, 12/01953-9, 17/07335-9, 15/15822-1)

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. (11/23110-0, 12/50161-8, 14/07206-6)

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. (14/07206-6, 14/16646-0, 12/50161-8, 11/23110-0)

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. (11/23110-0, 14/16646-0, 14/07206-6)

TIROLI-CEPEDA, ANA O.; SERAPHIM, THIAGO V.; PINHEIRO, GLAUCIA M. S.; SOUTO, DENIO E. P.; KUBOTA, LAURO T.; BORGES, JULIO C.; BARBOSA, LEANDRO R. S.; RAMOS, CARLOS H. I.. Studies on the effect of the J-domain on the substrate binding domain (SBD) of Hsp70 using a chimeric human J-SBD polypeptide. International Journal of Biological Macromolecules, v. 124, p. 111-120, MAR 1 2019. (14/07206-6, 17/07335-9, 15/15822-1, 12/50161-8)