Investigation of the ADAM17 metalloproteinase regulation mechanism by the interaction with Trx-1

• Grant number: 14/06485-9
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: February 01, 2015
• End date: August 05, 2018
• Field of knowledge: Health Sciences - Medicine
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Adriana Franco Paes Leme
• Grantee: Rute Alves Pereira e Costa
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Associated research grant: 10/19278-0 - Study of regulation of ADAMs in oral cancer, AP.JP
• Associated scholarship(s): 16/01528-7 - The effect of Trx-1 overexpression on protein expression and redox state of cysteines by iodoTMT and TMT labeling, BE.EP.PD

Abstract

ADAM17 is a membrane metalloproteinase which has the main function to modulate cell surface signaling events by release of surface protein ectodomains, rendering them solubility to interact with cellular receptors. Several studies have shown the modulation of ADAM17 activity and our group recently identified Trx-1 as interaction partner and suggested that overexpression of Trx-1 can be one of the pathways regulating ADAM17. However, this regulatory mechanism has not been elucidated yet. It is not known whether this regulation may be via (1) directly interaction of Trx-1 with ADAM17, (2) via conversion of reactive oxygen species (ROS) by Trx-1, since it has been proposed that ROS act in the activation of ADAM17, or (3) by indirect Trx-1 signaling on other targets or ligands. Thus, the goal of this project is to investigate these three possible mechanisms and for that, (1) we will mutate Trx-1, as well as use synthetic peptides of the interface interaction based on the published model (Aragão et al., 2012) to evaluate the effect on interaction and activity of ADAM17, (2) determine ROS by quantifying the O2 and H2O2 under Trx-1 overexpression conditions with the mutated C32 and C35 catalytic site, to assess ROS availability and thereby ADAM17 regulating activity. To answer the third hypothesis we will perform immunoprecipitation with Trx-1 mutated C32 and C35 catalytic site, following by identification and relative quantification of interaction partners for quantitative proteomics using SILAC, as well as their levels of phosphorylation. In addition, will access the redox state of the interaction partners of Trx-1 using the labeling of cysteines with ICAT. The interaction partners identified will be analyzed regarding their function and ability to interfere in the activity of ADAM17. We expect that this study can contribute to the elucidation of the ADAM17 regulation mechanism via Trx-1. (AU)

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)

GRANATO, DANIELA C.; P. E COSTA, RUTE A.; KAWAHARA, REBECA; YOKOO, SAMI; ARAGAO, ANNELIZE Z.; DOMINGUES, ROMENIA R.; PAULETTI, BIANCA A.; HONORATO, RODRIGO V.; FATTORI, JULIANA; FIGUEIRA, ANA CAROLINA M.; et al. Thioredoxin-1 Negatively Modulates ADAM17 Activity Through Direct Binding and Indirect Reductive Activity. Antioxidants & Redox Signaling, v. 29, n. 8, FEB 2018. (16/01528-7, 09/54067-3, 14/06485-9, 14/23888-0, 11/02267-9, 10/19278-0)

GRANATO, DANIELA C.; NEVES, LEANDRO X.; TRINO, LUCIANA D.; CARNIELLI, CAROLINA M.; LOPES, ARIANE F. B.; YOKOO, SAMI; PAULETTI, BIANCA A.; DOMINGUES, ROMENIA R.; SA, JAMILE O.; PERSINOTI, GABRIELLA; et al. Meta-omics analysis indicates the saliva microbiome and its proteins associated with the prognosis of oral cancer patients. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1869, n. 8, AUG 2021. (18/18496-6, 18/02180-0, 16/07846-0, 19/18751-9, 19/21815-9, 09/54067-3, 18/11958-4, 15/50590-4, 18/12194-8, 18/15728-3, 14/06485-9, 18/15535-0, 10/19278-0)

E COSTA, RUTE A. P.; GRANATO, DANIELA C.; TRINO, LUCIANA D.; YOKOO, SAMI; CARNIELLI, CAROLINA M.; KAWAHARA, REBECA; DOMINGUES, ROMENIA R.; PAULETTI, BIANCA ALVES; NEVES, LEANDRO XAVIER; SANTANA, ALINE G.; et al. ADAM17 cytoplasmic domain modulates Thioredoxin-1 conformation and activity. REDOX BIOLOGY, v. 37, OCT 2020. (16/01528-7, 18/12194-8, 19/18751-9, 18/18496-6, 09/54067-3, 18/15535-0, 14/23888-0, 16/07846-0, 16/24664-3, 14/06485-9, 10/19278-0)

RIVERA, CESAR; OLIVEIRA, ANA KARINA; PEREIRA COSTA, RUTE ALVES; DE ROSSI, TATIANE; PAES LEME, ADRIANA FRANCO. Prognostic biomarkers in oral squamous cell carcinoma: A systematic review. Oral Oncology, v. 72, p. 38-47, SEP 2017. (16/07846-0, 14/06485-9, 15/12431-1)