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Cysteine residues mediate high‐affinity binding of thioredoxin to ASK1

SYSNO ASEP	0465069
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Cysteine residues mediate high‐affinity binding of thioredoxin to ASK1
Author(s)	Kylarová, Salome (FGU-C) Košek, Dalibor (FGU-C)_{ORCID, RID} Petrvalská, Olivia (FGU-C)_{RID, ORCID, SAI} Pšenáková, Katarína (FGU-C)_{RID, ORCID} Man, Petr (MBU-M)_{RID, ORCID} Večeř, J. (CZ) Herman, P. (CZ) Obšilová, Veronika (FGU-C)_{RID, ORCID, SAI} Obšil, Tomáš (FGU-C)_{RID, ORCID}
Source Title	FEBS Journal - ISSN 1742-464X Roč. 283, č. 20 (2016), s. 3821-3838
Number of pages	18 s.
Language	eng - English
Country	GB - United Kingdom
Keywords	ASK 1 ; cysteine ; disulfide bond ; mass spectrometry ; TRX
Subject RIV	CE - Biochemistry
Subject RIV - cooperation	Institute of Microbiology - Microbiology, Virology
R&D Projects	GA14-10061S GA ČR - Czech Science Foundation (CSF)
	ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
Institutional support	FGU-C - RVO:67985823 ; MBU-M - RVO:61388971
UT WOS	000388284400011
EID SCOPUS	84988328603
DOI	10.1111/febs.13893
Annotation	Apoptosis signal-regulating kinase 1 (ASK1, MAP3K5) activates p38 mitogen-activated protein kinase and the c-Jun N-terminal kinase in response to proinflammatory and stress signals. In nonstress conditions, ASK1 is inhibited by association with thioredoxin (TRX) which binds to the TRX-binding domain (ASK1-TBD) at the N terminus of ASK1. TRX dissociates in response to oxidative stress allowing the ASK1 activation. However, the molecular basis for the ASK1:TRX1 complex dissociation is still not fully understood. Here, the role of cysteine residues on the interaction between TRX1 and ASK1-TBD in both reducing and oxidizing conditions was investigated. We show that from the two catalytic cysteines of TRX1 the residue C32 is responsible for the high-affinity binding of TRX1 to ASK1-TBD in reducing conditions. The disulfide bond formation between C32 and C35 within the active site of TRX1 is the main factor responsible for the TRX1 dissociation upon its oxidation as the formation of the second disulfide bond between noncatalytic cysteines C62 and C69 did not have any additional effect. ASK1-TBD contains seven conserved cysteine residues which differ in solvent accessibility with the residue C250 being the only cysteine which is both solvent exposed and essential for TRX1 binding in reducing conditions. Furthermore, our data show that the catalytic site of TRX1 interacts with ASK1-TBD region containing cysteine C200 and that the oxidative stress induces intramolecular disulfide bond formation within ASK1-TBD and affects its structure in regions directly involved and/or important for TRX1 binding.
Workplace	Institute of Physiology
Contact	Lucie Trajhanová, lucie.trajhanova@fgu.cas.cz, Tel.: 241 062 400
Year of Publishing	2017

Number of the records: 1