Universität zu Köln

Granato, Daniela C., P. e Costa, Rute A., Kawahara, Rebeca ORCID: 0000-0002-0292-2637, Yokoo, Sami, Aragao, Annelize Z., Domingues, Romenia R., Pauletti, Bianca A., Honorato, Rodrigo V., Fattori, Juliana ORCID: 0000-0002-5865-0183, Figueira, Ana Carolina M., Oliveira, Paulo S. L., Consonni, Silvio R., Fernandes, Denise, Laurindo, Francisco, Hansen, Hinrich P. and Leme, Adriana F. Paes (2018). Thioredoxin-1 Negatively Modulates ADAM17 Activity Through Direct Binding and Indirect Reductive Activity. Antioxid. Redox Signal., 29 (8). S. 717 - 735. NEW ROCHELLE: MARY ANN LIEBERT, INC. ISSN 1557-7716

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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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Granato, Daniela C. UNSPECIFIED UNSPECIFIED UNSPECIFIED P. e Costa, Rute A. UNSPECIFIED UNSPECIFIED UNSPECIFIED Kawahara, Rebeca UNSPECIFIED orcid.org/0000-0002-0292-2637 UNSPECIFIED Yokoo, Sami UNSPECIFIED UNSPECIFIED UNSPECIFIED Aragao, Annelize Z. UNSPECIFIED UNSPECIFIED UNSPECIFIED Domingues, Romenia R. UNSPECIFIED UNSPECIFIED UNSPECIFIED Pauletti, Bianca A. UNSPECIFIED UNSPECIFIED UNSPECIFIED Honorato, Rodrigo V. UNSPECIFIED UNSPECIFIED UNSPECIFIED Fattori, Juliana UNSPECIFIED orcid.org/0000-0002-5865-0183 UNSPECIFIED Figueira, Ana Carolina M. UNSPECIFIED UNSPECIFIED UNSPECIFIED Oliveira, Paulo S. L. UNSPECIFIED UNSPECIFIED UNSPECIFIED Consonni, Silvio R. UNSPECIFIED UNSPECIFIED UNSPECIFIED Fernandes, Denise UNSPECIFIED UNSPECIFIED UNSPECIFIED Laurindo, Francisco UNSPECIFIED UNSPECIFIED UNSPECIFIED Hansen, Hinrich P. UNSPECIFIED UNSPECIFIED UNSPECIFIED Leme, Adriana F. Paes UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-195494
DOI:	10.1089/ars.2017.7297
Journal or Publication Title:	Antioxid. Redox Signal.
Volume:	29
Number:	8
Page Range:	S. 717 - 735
Date:	2018
Publisher:	MARY ANN LIEBERT, INC
Place of Publication:	NEW ROCHELLE
ISSN:	1557-7716
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language NECROSIS-FACTOR-ALPHA; PROTEIN-DISULFIDE ISOMERASE; MAMMALIAN THIOREDOXIN; METALLOPROTEASE 17; SHEDDING ACTIVITY; MOLECULAR SWITCH; DISINTEGRIN; MECHANISM; CELLS; INFLAMMATION Multiple languages Biochemistry & Molecular Biology; Endocrinology & Metabolism Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/19549