| Full text | |
| Author(s): |
Pinheiro, Glaucia M. S.
[1]
;
Amorim, Gisele C.
[2, 3]
;
Iqbal, Anwar
[2, 4]
;
Almeida, Fabio C. L.
[5, 2]
;
Ramosa, C. H. I.
[5, 1]
Total Authors: 5
|
| Affiliation: | [1] Univ Campinas UNICAMP, Inst Chem, Campinas, SP - Brazil
[2] Univ Fed Rio de Janeiro, Inst Med Biochem & Nucl Struct Biol & Bioimaging, Rio De Janeiro, RJ - Brazil
[3] Univ Fed Rio de Janeiro, Nucleo Multidiscipliner Pesquisa Biol, Campus Duque Caxias, Rio De Janeiro, RJ - Brazil
[4] Gomal Univ, Inst Chem Sci, Dera Ismail Khan, KP - Pakistan
[5] Inst Nacl Ciencia & Tecnol Biol Estrutural & Bioi, Rio De Janeiro - Brazil
Total Affiliations: 5
|
| Document type: | Journal article |
| Source: | Archives of Biochemistry and Biophysics; v. 669, p. 71-79, JUL 15 2019. |
| Web of Science Citations: | 1 |
| Abstract | |
J-domain/Hsp40 proteins cooperate in aiding with folding in the cell by binding partially folded client proteins and delivering them to be folded by Hsp70. The delivery occurs concomitantly to the stimulation of the ATPase activity of Hsp70 via the N-terminally located J-domain. Although several lines of investigation have been used to study J-domain proteins, the presence of highly flexible domains (G/F- and G/M-rich) hold up obtaining a detailed full-length structure. In this work, we present the high-resolution structure of the J-domain and the N-terminal part of the G/F domain of Sis1, solved by NMR, and used chemical-shift perturbation approaches to further study the structure/function relationship of the Sis1/Hsp70 interaction. When the J-domain was compared to the full-length protein and to a G/M domain deletion mutant, an internal interaction patch formed by hydrophobic and positively charged residues (V2, D9, R27, T39, F52 and R73) was identified. Curiously, the same patch is protected by internal contacts in the full-length protein and, in combination with the loop containing the conserved HPD motif, participates in the interaction with Hsp70. Combined, these results suggest that the J-domain in the full-length Sis1 is in a transient intermediate conformation, in which its interacting patch is protected and, at the same time, also in a favorable condition to bind Hsp70, facilitating the interaction between the two proteins. Finally, 1D NMR experiments showed that the addition of ATP is followed by the disruption of the J-domain/Hsp70 complex, a necessary step for aiding the folding of the client protein. (AU) | |
| FAPESP's process: | 17/01074-9 - HSP90 interaction with other proteins and cofactors |
| Grantee: | Glaucia Melina Squizato Pinheiro de Castro |
| Support type: | Scholarships in Brazil - Post-Doctorate |
| FAPESP's process: | 18/11948-9 - Interaction mechanism between the co-chaperone J-domain protein with their substrates and the chaperone Hsp70 |
| Grantee: | Glaucia Melina Squizato Pinheiro de Castro |
| Support type: | Scholarships in Brazil - Post-Doctorate |
| FAPESP's process: | 12/50161-8 - Study of the structure and function of the Hsp90 chaperone with emphasis on its role in cellular homeostasis |
| Grantee: | Carlos Henrique Inacio Ramos |
| Support type: | Research Projects - Thematic Grants |
| FAPESP's process: | 17/26131-5 - The chaperome: study of the relationship of the structure of its components and the maintenance of proteostasis |
| Grantee: | Carlos Henrique Inacio Ramos |
| Support type: | Research Projects - Thematic Grants |