Investigation of the ADAM17 metalloproteinase regulation mechanism by the interact...
| Full text | |
| Author(s): Show less - |
Granato, Daniela C.
[1]
;
P. e Costa, Rute A.
[1]
;
Kawahara, Rebeca
[1]
;
Yokoo, Sami
[1]
;
Aragao, Annelize Z.
[1]
;
Domingues, Romenia R.
[1]
;
Pauletti, Bianca A.
[1]
;
Honorato, Rodrigo V.
[1]
;
Fattori, Juliana
[1]
;
Figueira, Ana Carolina M.
[1]
;
Oliveira, Paulo S. L.
[1]
;
Consonni, Silvio R.
[1]
;
Fernandes, Denise
[2]
;
Laurindo, Francisco
[2]
;
Hansen, Hinrich P.
[3]
;
Leme, Adriana F. Paes
[1]
Total Authors: 16
|
| Affiliation: | [1] CNPEM, LNBio, Lab Nacl Biociencias, BR-13083970 Campinas, SP - Brazil
[2] Univ Sao Paulo, Fac Med, Inst Coracao, Sao Paulo - Brazil
[3] Univ Hosp Cologne, Dept Internal Med 1, Cologne - Germany
Total Affiliations: 3
|
| Document type: | Journal article |
| Source: | Antioxidants & Redox Signaling; v. 29, n. 8 FEB 2018. |
| Web of Science Citations: | 0 |
| Abstract | |
Aims: A disintegrin and metalloprotease 17 (ADAM17) modulates signaling events by releasing surface protein ectodomains such as TNFa and the EGFR-ligands. We have previously characterized cytoplasmic thioredoxin-1 (Trx-1) as a partner of ADAM17 cytoplasmic domain. Still, the mechanism of ADAM17 regulation by Trx-1 is unknown, and it has become of paramount importance to assess the degree of influence that Trx-1 has on metalloproteinase ADAM17. Results: Combining discovery and targeted proteomic approaches, we uncovered that Trx-1 negatively regulates ADAM17 by direct and indirect effect. We performed cell-based assays with synthetic peptides and site-directed mutagenesis, and we demonstrated that the interaction interface of Trx-1 and ADAM17 is important for the negative regulation of ADAM17 activity. However, both Trx-1(K72A) and catalytic site mutant Trx-1(C32/35S) rescued ADAM17 activity, although the interaction with Trx-1(C32/35S) was unaffected, suggesting an indirect effect of Trx-1. We confirmed that the Trx-1(C32/35S) mutant showed diminished reductive capacity, explaining this indirect effect on increasing ADAM17 activity through oxidant levels. Interestingly, Trx-1(K72A) mutant showed similar oxidant levels to Trx-1(C32/35S), even though its catalytic site was preserved. We further demonstrated that the general reactive oxygen species inhibitor, Nacetylcysteine (NAC), maintained the regulation of ADAM17 dependent of Trx-1 reductase activity levels; whereas the electron transport chain modulator, rotenone, abolished Trx-1 effect on ADAM17 activity. Innovation: We show for the first time that the mechanism of ADAM17 regulation, Trx-1 dependent, can be by direct interaction and indirect effect, bringing new insights into the cross-talk between isomerases and mammalian metalloproteinases. Conclusion: This unexpected Trx-1(K72A) behavior was due to more dimer formation and, consequently, the reduction of its Trx-1 reductase activity, evaluated through dimer verification, by gel filtration and mass spectrometry analysis. (AU) | |
| FAPESP's process: | 16/01528-7 - The effect of Trx-1 overexpression on protein expression and redox state of cysteines by iodoTMT and TMT labeling |
| Grantee: | Rute Alves Pereira e Costa |
| Support type: | Scholarships abroad - Research Internship - Post-doctor |
| FAPESP's process: | 09/54067-3 - Acquisition of a mass spectrometer coupled to a liquid chromatography system for increasing the capacity to meet the needs of users and for making new technologies available in the Laboratory of Mass Spectrometry |
| Grantee: | Adriana Franco Paes Leme |
| Support type: | Multi-user Equipment Program |
| FAPESP's process: | 14/06485-9 - Investigation of the ADAM17 metalloproteinase regulation mechanism by the interaction with Trx-1 |
| Grantee: | Rute Alves Pereira e Costa |
| Support type: | Scholarships in Brazil - Post-Doctorate |
| FAPESP's process: | 14/23888-0 - Studies on the activation mechanisms of MMP-2 and ADAM17: identification of regulatory proteins, oxidant production pathways, epigenetics and proteolytic targets |
| Grantee: | Raquel Fernanda Gerlach |
| Support type: | Research Projects - Thematic Grants |
| FAPESP's process: | 11/02267-9 - Identifying ADAM17 partners and mapping the domains responsible for its interaction with other proteins |
| Grantee: | Daniela Campos Granato |
| Support type: | Scholarships in Brazil - Post-Doctorate |
| FAPESP's process: | 10/19278-0 - Study of regulation of ADAMs in oral cancer |
| Grantee: | Adriana Franco Paes Leme |
| Support type: | Research Grants - Young Investigators Grants |