Dataset Information

ADAM17 cytoplasmic domain modulates Thioredoxin-1 conformation and activity

ABSTRACT: We have demonstrated that site-directed mutagenesis in ADAM17 (ADAM17cytoF730A) also disrupts the interacting interface with Trx-1, resulting in a decrease of Trx-1 reductive capacity and activity. One of the mechanisms that explain this effect might be that ADAM17cyto favors Trx-1 monomerization state - the active state - by forming a disulfide bond between Cys824 at the C-terminal of ADAM17cyto with the Cys73 of Trx-1, which is involved in the dimerization site. Moreover, we observed that ADAM17 overexpressing or knockdown cells favors or not the monomeric state of Trx-1, respectively. As a result, there is a decrease of oxidant levels and ADAM17 sheddase activity, and an increase in the reduced cysteine-containing peptides in intracellular proteins in ADAM17cyto overexpressing cells. In summary, we propose that ADAM17 is able to modulate Trx-1 conformation affecting its activity and intracellular redox state.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

SUBMITTER: Adriana Franco Paes Leme

LAB HEAD: Adriana Franco Paes Leme

PROVIDER: PXD015431 | Pride | 2020-10-19

REPOSITORIES: Pride

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ADAM17 cytoplasmic domain modulates Thioredoxin-1 conformation and activity.

E Costa Rute A P RAP Granato Daniela C DC Trino Luciana D LD Yokoo Sami S Carnielli Carolina M CM Kawahara Rebeca R Domingues Romênia R RR Pauletti Bianca Alves BA Neves Leandro Xavier LX Santana Aline G AG Paulo Joao A JA Aragão Annelize Z B AZB Heleno Batista Fernanda Aparecida FA Migliorini Figueira Ana Carolina AC Laurindo Francisco R M FRM Fernandes Denise D Hansen Hinrich P HP Squina Fabio F Gygi Steven P SP Paes Leme Adriana F AF

Redox biology 20200924

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 a ...[more]

PMID: 33011677

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Project description:Mutations in the gene encoding Cu-Zn superoxide dismutase 1 (SOD1) cause a subset of familial amyotrophic lateral sclerosis (fALS) cases. A shared effect of these mutations is that SOD1, which is normally a stable dimer, dissociates into toxic monomers that seed toxic aggregates. Considerable research effort has been devoted to developing compounds that stabilize the dimer of fALS SOD1 variants, but unfortunately, this has not yet resulted in a treatment. We hypothesized that cyclic thiosulfinate cross-linkers, which selectively target a rare, two cysteine-containing motif, can stabilize fALS-causing SOD1 variants in vivo. We created a library of chemically diverse cyclic thiosulfinates and determined structure-cross-linking-activity relationships. A pre-lead compound, “S-XL6,” was selected based upon its cross-linking rate and drug-like properties. Co-crystallographic structure clearly establishes the binding of S-XL6 at Cys 111 bridging the monomers and stabilizing the SOD1 dimer. Biophysical studies reveal that the degree of stabilization afforded by S-XL6 (up to 24 °C) is unprecedented for fALS, and to our knowledge, for any protein target of any kinetic stabilizer. Gene silencing and protein degrading therapeutic approaches require careful dose titration to balance the benefit of diminished fALS SOD1 expression with the toxic loss-of enzymatic function. We show that S-XL6 does not share this liability because it rescues the activity of fALS SOD1 variants. No pharmacological agent has been proven to bind to SOD1 in vivo. Here, using a fALS mouse model, we demonstrate oral bioavailability; rapid engagement of SOD1G93A by S-XL6 that increases SOD1G93A’s in vivo half-life; and that S-XL6 crosses the blood-brain barrier. S-XL6 demonstrated a degree of selectivity by avoiding off-target binding to plasma proteins. Taken together, our results indicate that cyclic thiosulfinate-mediated SOD1 stabilization should receive further attention as a potential therapeutic approach for fALS.

			Action	DRS
	09jun_Trx_D_DG_1.mgf	Mgf
	09jun_Trx_D_DG_1.msf	Msf
	09jun_Trx_D_DG_1.raw	Raw
	09jun_Trx_D_DG_2.mgf	Mgf
	09jun_Trx_D_DG_2.msf	Msf