ADAM17 cytoplasmic domain modulates Thioredoxin-1 conformation and activity

Rute A. P. e Costa; Daniela C. Granato; Luciana D Trino; Sami Yokoo; Carolina M. Carnielli; Rebeca Kawahara; Romênia R. Domingues; Bianca Alves Pauletti; Leandro Xavier Neves; Aline G. Santana; Joao A. Paulo; Annelize Z. B. Aragão; Fernanda Aparecida Heleno Batista; Ana Carolina Migliorini Figueira; Francisco R. M. Laurindo; Denise Fernandes; Hinrich P. Hansen; Fabio Squina; Steven P. Gygi; Adriana F. Paes Leme

doi:10.1016/j.redox.2020.101735

ADAM17 cytoplasmic domain modulates Thioredoxin-1 conformation and activity

Rute A. P. e Costa, Daniela C. Granato, Luciana D Trino, Sami Yokoo, Carolina M. Carnielli, Rebeca Kawahara, Romênia R. Domingues, Bianca Alves Pauletti, Leandro Xavier Neves, Aline G. Santana, Joao A. Paulo, Annelize Z. B. Aragão, Fernanda Aparecida Heleno Batista, Ana Carolina Migliorini Figueira, Francisco R. M. Laurindo, Denise Fernandes, Hinrich P. Hansen, Fabio Squina, Steven P. Gygi, Adriana F. Paes Leme^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

2 Citations (Scopus)

5 Downloads (Pure)

Abstract

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 (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 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.

Original language	English
Article number	101735
Pages (from-to)	1-16
Number of pages	16
Journal	Redox Biology
Volume	37
DOIs	https://doi.org/10.1016/j.redox.2020.101735
Publication status	Published - Oct 2020
Externally published	Yes

Keywords

ADAM17 Protein
Cysteine/metabolism
HEK293 Cells
Humans
Molecular Conformation
Oxidation-Reduction
Sulfhydryl Compounds
Thioredoxins/genetics
iodoTMT
Redox signaling
Thioredoxin-1
ADAM17
Dimerization
Mass spectrometry

Access to Document

10.1016/j.redox.2020.101735

Publisher version (open access)Final published version, 8.11 MBLicence: CC BY-NC-ND

Cite this

e Costa, R. A. P., Granato, D. C., Trino, L. D., Yokoo, S., Carnielli, C. M., Kawahara, R., Domingues, R. R., Pauletti, B. A., Neves, L. X., Santana, A. G., Paulo, J. A., Aragão, A. Z. B., Heleno Batista, F. A., Migliorini Figueira, A. C., Laurindo, F. R. M., Fernandes, D., Hansen, H. P., Squina, F., Gygi, S. P., & Paes Leme, A. F. (2020). ADAM17 cytoplasmic domain modulates Thioredoxin-1 conformation and activity. Redox Biology, 37, 1-16. [101735]. https://doi.org/10.1016/j.redox.2020.101735

e Costa, Rute A. P. ; Granato, Daniela C. ; Trino, Luciana D ; Yokoo, Sami ; Carnielli, Carolina M. ; Kawahara, Rebeca ; Domingues, Romênia R. ; Pauletti, Bianca Alves ; Neves, Leandro Xavier ; Santana, Aline G. ; Paulo, Joao A. ; Aragão, Annelize Z. B. ; Heleno Batista, Fernanda Aparecida ; Migliorini Figueira, Ana Carolina ; Laurindo, Francisco R. M. ; Fernandes, Denise ; Hansen, Hinrich P. ; Squina, Fabio ; Gygi, Steven P. ; Paes Leme, Adriana F. / ADAM17 cytoplasmic domain modulates Thioredoxin-1 conformation and activity. In: Redox Biology. 2020 ; Vol. 37. pp. 1-16.

@article{f2b5bf1835fe485cb2ba8cffbd9664fe,

title = "ADAM17 cytoplasmic domain modulates Thioredoxin-1 conformation and activity",

abstract = "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.",

keywords = "ADAM17 Protein, Cysteine/metabolism, HEK293 Cells, Humans, Molecular Conformation, Oxidation-Reduction, Sulfhydryl Compounds, Thioredoxins/genetics, iodoTMT, Redox signaling, Thioredoxin-1, ADAM17, Dimerization, Mass spectrometry",

author = "{e Costa}, {Rute A. P.} and Granato, {Daniela C.} and Trino, {Luciana D} and Sami Yokoo and Carnielli, {Carolina M.} and Rebeca Kawahara and Domingues, {Rom{\^e}nia R.} and Pauletti, {Bianca Alves} and Neves, {Leandro Xavier} and Santana, {Aline G.} and Paulo, {Joao A.} and Arag{\~a}o, {Annelize Z. B.} and {Heleno Batista}, {Fernanda Aparecida} and {Migliorini Figueira}, {Ana Carolina} and Laurindo, {Francisco R. M.} and Denise Fernandes and Hansen, {Hinrich P.} and Fabio Squina and Gygi, {Steven P.} and {Paes Leme}, {Adriana F.}",

note = "Copyright {\textcopyright} 2020. Published by Elsevier B.V.",

year = "2020",

month = oct,

doi = "10.1016/j.redox.2020.101735",

language = "English",

volume = "37",

pages = "1--16",

journal = "Redox Biology",

issn = "2213-2317",

publisher = "Elsevier",

}

e Costa, RAP, Granato, DC, Trino, LD, Yokoo, S, Carnielli, CM, Kawahara, R, Domingues, RR, Pauletti, BA, Neves, LX, Santana, AG, Paulo, JA, Aragão, AZB, Heleno Batista, FA, Migliorini Figueira, AC, Laurindo, FRM, Fernandes, D, Hansen, HP, Squina, F, Gygi, SP & Paes Leme, AF 2020, 'ADAM17 cytoplasmic domain modulates Thioredoxin-1 conformation and activity', Redox Biology, vol. 37, 101735, pp. 1-16. https://doi.org/10.1016/j.redox.2020.101735

ADAM17 cytoplasmic domain modulates Thioredoxin-1 conformation and activity. / e Costa, Rute A. P.; Granato, Daniela C.; Trino, Luciana D; Yokoo, Sami; Carnielli, Carolina M.; Kawahara, Rebeca; Domingues, Romênia R.; Pauletti, Bianca Alves; Neves, Leandro Xavier; Santana, Aline G.; Paulo, Joao A.; Aragão, Annelize Z. B.; Heleno Batista, Fernanda Aparecida; Migliorini Figueira, Ana Carolina; Laurindo, Francisco R. M.; Fernandes, Denise; Hansen, Hinrich P.; Squina, Fabio; Gygi, Steven P.; Paes Leme, Adriana F.

In: Redox Biology, Vol. 37, 101735, 10.2020, p. 1-16.

Research output: Contribution to journal › Article › peer-review

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.

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 -

ADAM17 cytoplasmic domain modulates Thioredoxin-1 conformation and activity

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Cite this