| Texto completo | |
| Autor(es): |
Tiroli-Cepeda, Ana O.
[1]
;
Seraphim, Thiago V.
[1]
;
Pinheiro, Glaucia M. S.
[1]
;
Souto, Denio E. P.
[1]
;
Kubota, Lauro T.
[1, 2]
;
Borges, Julio C.
[3]
;
Barbosa, Leandro R. S.
[4]
;
Ramos, Carlos H. I.
[1, 5]
Número total de Autores: 8
|
| Afiliação do(s) autor(es): | [1] Univ Estadual Campinas, BR-13083970 Campinas, SP - Brazil
[2] INCTBioanalitica, Campinas, SP - Brazil
[3] Univ Sao Paulo, Sao Carlos Inst Chem, BR-13560970 Sao Carlos, SP - Brazil
[4] Univ Sao Paulo, Inst Phys, Sao Paulo, SP - Brazil
[5] INBEB, Londrina - Brazil
Número total de Afiliações: 5
|
| Tipo de documento: | Artigo Científico |
| Fonte: | International Journal of Biological Macromolecules; v. 124, p. 111-120, MAR 1 2019. |
| Citações Web of Science: | 0 |
| Resumo | |
DnaJ/Hsp40 chaperones deliver unfolded proteins and stimulate the ATPase activity of DnaK/Hsp70 via their J-domain. However, the interaction is transient, creating a challenge for detailed analysis. We investigated whether it would be possible to gain further understanding of this interaction by engineering a chimeric polypeptide where the J-domain of Hsp40 was covalently attached to the substrate binding domain (SBD) of Hsp70 by a flexible linker. The rationale is to increase the proximity between the interacting partners to promote their natural interaction and facilitate the characterization of the interaction. The resulting chimera, termed J-SBD, was properly folded and had properties not present in the full-length Hsp70 or in the SBD alone, for instance a higher protective effect against aggregation and being a monomer. Substrate binding also appear to exceed that of SBD alone as revealed by a decreased binding to bis-ANS, a probe for hydrophobic patches. This hypothesis is supported by the structural model created by small angle X-ray scattering, suggesting that the lid subdomain (SBD alpha) is partially opened in the J-SBD. Collectively, our results suggest a model in which J-domain binding may shift the Hsp70 equilibrium towards the monomer state, exposing hydrophobic sites prone to substrate accommodation. (C) 2018 Elsevier B.V. All rights reserved. (AU) | |
| Processo FAPESP: | 14/07206-6 - Estudos da HSP70 mitocondrial de humanos e de protozoários: abordagem estrutural e funcional |
| Beneficiário: | Julio Cesar Borges |
| Modalidade de apoio: | Auxílio à Pesquisa - Regular |
| Processo FAPESP: | 17/07335-9 - Estudos das isoformas da HSP70 humana residentes no citoplasma e mitocôndria e de seus oligômeros de alta massa molecular: interação com co-chaperonas e proteínas clientes |
| Beneficiário: | Julio Cesar Borges |
| Modalidade de apoio: | Auxílio à Pesquisa - Regular |
| Processo FAPESP: | 15/15822-1 - Estudo das propriedades físico-químicas e estruturais de fármacos e líquidos iônicos com sistemas de relevância biológica |
| Beneficiário: | Leandro Ramos Souza Barbosa |
| Modalidade de apoio: | Auxílio à Pesquisa - Regular |
| Processo FAPESP: | 12/50161-8 - Estudo da estrutura e função da chaperona Hsp90 com ênfase no seu papel em homeostase celular |
| Beneficiário: | Carlos Henrique Inacio Ramos |
| Modalidade de apoio: | Auxílio à Pesquisa - Temático |