Thioredoxin-1 Negatively Modulates ADAM17 Activity Through Direct Binding and Indirect Reductive Activity

Granato, Daniela Campos; Costa, Rute Alves Pereira e; Kawahara, Rebeca; Yokoo, Sami; Aragão, Annelize Zambon Barbosa; Domingues, Romênia R.; Pauletti, Bianca Alves; Honorato, Rodrigo Vargas; Fattori, Juliana; Figueira, Ana Carolina Migliorini; Oliveira, Paulo Sérgio Lopes de; Consonni, Sílvio Roberto; Fernandes, Denise de Castro; Laurindo, Francisco Rafael Martins; Hansen, Hinrich P.; Leme, Adriana Franco Paes

doi:10.1089/ars.2017.7297

Cited by 10 publications

(18 citation statements)

References 41 publications

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“…Also, these results reinforced that ADAM17cyto WT directly and positively affected the Trx-1 redox activity. Besides, it corroborated with previous results showing that Trx-1 and reactive oxygen species modulated ADAM17 activity [ 28 , 40 ].…”

Section: Discussionsupporting

confidence: 91%

“…Also to evaluate whether ADAM17cyto WT modulated Trx-1 cysteine reactivity by interfering with the oxidation-reduction state of the catalytic site (Cys 32 -Cys 35 ) or the dimeric state (Cys 73 -Cys 73 ), the cysteine reactivity assay was performed with ADAM17cyto WT and Trx-1 mutants (Trx-1 K72A and Trx-1 C32/35S ). As expected, the activity of catalytic site mutant was not changed, but we showed that Trx-1 K72A , a catalytically inactive mutant due to its dimeric form [ 28 ], could have its cysteine reactivity increased by the incubation with ADAM17cyto WT , suggesting the ability of ADAM17cyto WT in the increase of Trx-1 activity acting upon the site of dimerization (Cys 73 -Cys 73 ) ( Fig. 2 D).…”

Section: Resultssupporting

confidence: 71%

“…Considering the evidences of the dimer and monomer forms of Trx-1 [ 24 , 28 ], we studied whether ADAM17 contributes with these states according to the described experimental design ( Fig. 1 ).…”

Section: Resultsmentioning

confidence: 99%

“…In the in silico ADAM17cyto and Trx-1 complex modeling, the residue F730 of ADAM17 was previously shown to be spatially located near the amino acid residue K72 of Trx-1, residue that was also shown to contribute to the binding interface and modeling of Trx-1 dimers (Cys 73 -Cys 73 ’ disulfide bond in direct vicinity to K72) [ 27 , 28 ]. Therefore, we have interrogated if the interaction with ADAM17cyto WT could modulate or influence the oligomerization state of Trx-1 and, thus, affect the oxidation/reduction state of the cell.…”

Section: Discussionmentioning

confidence: 99%

“…This interaction is in the opposite location of the catalytic site of Trx-1. A recent work from our group [ 28 ] has shown that ADAM17/Trx-1-interacting interface was destabilized by a mutation in Trx-1 (Trx-1 K72A ), favoring the Cys 73 -Cys 73 ’ homodimeric form of Trx-1, which is the inactive state of Trx-1. Therefore, we hypothesized whether the interface of ADAM17/Trx-1 may also play a role in the Trx-1 activity.…”

Section: Introductionmentioning

confidence: 99%

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ADAM17 cytoplasmic domain modulates Thioredoxin-1 conformation and activity

et al. 2020

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The activity of Thioredoxin-1 (Trx-1) is adjusted by the balance of its monomeric, active and its dimeric, inactive state. The regulation of this balance is not completely understood. We have previously shown that the cytoplasmic domain of the transmembrane protein A D isintegrin A nd M etalloprotease 17 (ADAM17cyto) binds to Thioredoxin-1 (Trx-1) and the destabilization of this interaction favors the dimeric state of Trx-1. Here, we investigate whether ADAM17 plays a role in the conformation and activation of Trx-1. We found that disrupting the interacting interface with Trx-1 by a site-directed mutagenesis in ADAM17 (ADAM17cyto F730A ) caused a decrease of Trx-1 reductive capacity and activity. Moreover, we observed that ADAM17 overexpressing cells favor the monomeric state of Trx-1 while knockdown cells do not. As a result, there is a decrease of cell oxidant levels and ADAM17 sheddase activity and an increase in the reduced cysteine-containing peptides in intracellular proteins in ADAM17cyto overexpressing cells. A mechanistic explanation that ADAM17cyto favors the monomeric, active state of Trx-1 is the formation of a disulfide bond between Cys 824 at the C-terminal of ADAM17cyto with the Cys 73 of Trx-1, which is involved in the dimerization site of Trx-1. In summary, we propose that ADAM17 is able to modulate Trx-1 conformation affecting its activity and intracellular redox state, bringing up a novel possibility for positive regulation of thiol isomerase activity in the cell by mammalian metalloproteinases.

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Section: Discussionsupporting

confidence: 91%

Section: Resultssupporting

confidence: 71%