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dc.contributor.authorHughes, Mark M.
dc.contributor.authorLavrencic, Peter
dc.contributor.authorColl, Rebecca C.
dc.contributor.authorVe, Thomas
dc.contributor.authorRyan, Dylan G.
dc.contributor.authorWilliams, Niamh C.
dc.contributor.authorMenon, Deepthi
dc.contributor.authorMansell, Ashley
dc.contributor.authorBoard, Philip G.
dc.contributor.authorMobli, Mehdi
dc.contributor.authorKobe, Bostjan
dc.contributor.authorO'Neill, Luke A. J.
dc.date.accessioned2017-09-22T04:28:38Z
dc.date.available2017-09-22T04:28:38Z
dc.date.issued2017
dc.identifier.issn0027-8424en_US
dc.identifier.doi10.1073/pnas.1701868114en_US
dc.identifier.urihttp://hdl.handle.net/10072/347156
dc.description.abstractMyD88 adaptor-like (MAL) is a critical protein in innate immunity, involved in signaling by several Toll-like receptors (TLRs), key pattern recognition receptors (PRRs). Crystal structures of MAL revealed a nontypical Toll/interleukin-1 receptor (TIR)-domain fold stabilized by two disulfide bridges. We therefore undertook a structural and functional analysis of the role of reactive cysteine residues in the protein. Under reducing conditions, the cysteines do not form disulfides, but under oxidizing conditions they are highly amenable to modification. The solution structure of the reduced form of the MAL TIR domain, determined by NMR spectroscopy, reveals a remarkable structural rearrangement compared with the disulfide-bonded structure, which includes the relocation of a β-strand and repositioning of the functionally important “BB-loop” region to a location more typical for TIR domains. Redox measurements by NMR further reveal that C91 has the highest redox potential of all cysteines in MAL. Indeed, mass spectrometry revealed that C91 undergoes glutathionylation in macrophages activated with the TLR4 ligand lipopolysaccharide (LPS). The C91A mutation limits MAL glutathionylation and acts as a dominant negative, blocking the interaction of MAL with its downstream target MyD88. The H92P mutation mimics the dominant-negative effects of the C91A mutation, presumably by preventing C91 glutathionylation. The MAL C91A and H92P mutants also display diminished degradation and interaction with interleukin-1 receptor-associated kinase 4 (IRAK4). We conclude that in the cell, MAL is not disulfide-bonded and requires glutathionylation of C91 for signaling.en_US
dc.description.peerreviewedYesen_US
dc.languageEnglishen_US
dc.publisherNational Academy of Sciencesen_US
dc.relation.ispartofpagefromE6480en_US
dc.relation.ispartofpagetoE6489en_US
dc.relation.ispartofissue32en_US
dc.relation.ispartofjournalProceedings of the National Academy of Sciences of the United States of Americaen_US
dc.relation.ispartofvolume114en_US
dc.subject.fieldofresearchAnalytical Biochemistryen_US
dc.subject.fieldofresearchcode060101en_US
dc.titleSolution structure of the TLR adaptor MAL/TIRAP reveals an intact BB loop and supports MAL Cys91 glutathionylation for signalingen_US
dc.typeJournal articleen_US
dc.type.descriptionC1 - Peer Reviewed (HERDC)en_US
dc.type.codeC - Journal Articlesen_US
gro.hasfulltextNo Full Text


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