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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Identification of Novel Interaction between ADAM17 (a Disintegrin and Metalloprotease 17) and Thioredoxin-1

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Author(s):
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Aragao, Annelize Z. B. [1] ; Nogueira, Maria Luiza C. [1] ; Granato, Daniela C. [1] ; Simabuco, Fernando M. [1] ; Honorato, Rodrigo V. [1] ; Hoffman, Zaira [2] ; Yokoo, Sami [1] ; Laurindo, Francisco R. M. [3] ; Squina, Fabio M. [2] ; Zeri, Ana Carolina M. [1] ; Oliveira, Paulo S. L. [1] ; Sherman, Nicholas E. [4] ; Paes Leme, Adriana F. [1]
Total Authors: 13
Affiliation:
[1] CNPEM, Lab Espectrometria Massas, Lab Nacl Biociencias, LNBio, BR-13083970 Campinas, SP - Brazil
[2] CNPEM, Lab Nacl Ciencia & Tecnol Bioetanol, CTBE, BR-13083970 Campinas, SP - Brazil
[3] Univ Sao Paulo, Inst Coracao, BR-05403000 Sao Paulo - Brazil
[4] Univ Virginia, WM Keck Biomed Mass Spectrometry Lab, Charlottesville, VA 22908 - USA
Total Affiliations: 4
Document type: Journal article
Source: Journal of Biological Chemistry; v. 287, n. 51, p. 43071-43082, DEC 14 2012.
Web of Science Citations: 22
Abstract

ADAM17, which is also known as TNF alpha-converting enzyme, is the major sheddase for the EGF receptor ligands and is considered to be one of the main proteases responsible for the ectodomain shedding of surface proteins. How a membrane-anchored proteinase with an extracellular catalytic domain can be activated by inside-out regulation is not completely understood. We characterized thioredoxin-1 (Trx-1) as a partner of the ADAM17 cytoplasmic domain that could be involved in the regulation of ADAM17 activity. We induced the overexpression of the ADAM17 cytoplasmic domain in HEK293 cells, and ligands able to bind this domain were identified by MS after protein immunoprecipitation. Trx-1 was also validated as a ligand of the ADAM17 cytoplasmic domain and full-length ADAM17 recombinant proteins by immunoblotting, immunolocalization, and solid phase binding assay. In addition, using nuclear magnetic resonance, it was shown in vitro that the titration of the ADAM17 cytoplasmic domain promotes changes in the conformation of Trx-1. The MS analysis of the cross-linked complexes showed cross-linking between the two proteins by lysine residues. To further evaluate the functional role of Trx-1, we used a heparin-binding EGF shedding cell model and observed that the overexpression of Trx-1 in HEK293 cells could decrease the activity of ADAM17, activated by either phorbol 12-myristate 13-acetate or EGF. This study identifies Trx-1 as a novel interaction partner of the ADAM17 cytoplasmic domain and suggests that Trx-1 is a potential candidate that could be involved in ADAM17 activity regulation. (AU)

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: 09/18301-1 - Study of the regulation of recombinant ADAM-17 by determining phosphorylation sites of the cytoplasmic domain and its ligands in normal and cancer cells
Grantee:Annelize Zambon Barbosa Aragão
Support type: Scholarships in Brazil - Doctorate
FAPESP's process: 10/15675-5 - Effects of modulation of the membrane metalloproteinase ADAM-17 in the extracelular subproteome using oral cancer as a model
Grantee:Fernando Moreira Simabuco
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
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