TY - JOUR
T1 - ADAM17 cytoplasmic domain modulates Thioredoxin-1 conformation and activity
AU - e Costa, Rute A. P.
AU - Granato, Daniela C.
AU - Trino, Luciana D
AU - Yokoo, Sami
AU - Carnielli, Carolina M.
AU - Kawahara, Rebeca
AU - Domingues, Romênia R.
AU - Pauletti, Bianca Alves
AU - Neves, Leandro Xavier
AU - Santana, Aline G.
AU - Paulo, Joao A.
AU - Aragão, Annelize Z. B.
AU - Heleno Batista, Fernanda Aparecida
AU - Migliorini Figueira, Ana Carolina
AU - Laurindo, Francisco R. M.
AU - Fernandes, Denise
AU - Hansen, Hinrich P.
AU - Squina, Fabio
AU - Gygi, Steven P.
AU - Paes Leme, Adriana F.
N1 - Copyright © 2020. Published by Elsevier B.V.
PY - 2020/10
Y1 - 2020/10
N2 - 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 Disintegrin And Metalloprotease 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 (ADAM17cytoF730A) 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 Cys824 at the C-terminal of ADAM17cyto with the Cys73 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.
AB - 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 Disintegrin And Metalloprotease 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 (ADAM17cytoF730A) 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 Cys824 at the C-terminal of ADAM17cyto with the Cys73 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.
KW - ADAM17 Protein
KW - Cysteine/metabolism
KW - HEK293 Cells
KW - Humans
KW - Molecular Conformation
KW - Oxidation-Reduction
KW - Sulfhydryl Compounds
KW - Thioredoxins/genetics
KW - iodoTMT
KW - Redox signaling
KW - Thioredoxin-1
KW - ADAM17
KW - Dimerization
KW - Mass spectrometry
UR - http://www.scopus.com/inward/record.url?scp=85092324570&partnerID=8YFLogxK
U2 - 10.1016/j.redox.2020.101735
DO - 10.1016/j.redox.2020.101735
M3 - Article
C2 - 33011677
VL - 37
SP - 1
EP - 16
JO - Redox Biology
JF - Redox Biology
SN - 2213-2317
M1 - 101735
ER -