PDBsum entry 2ndh

Immune system

PDB id: 2ndh

Contents

Protein chain

143 a.a.

PDB id:

2ndh

Name:

Immune system

Title:

Nmr solution structure of mal/tirap tir domain (c116a)

Structure:

Toll/interleukin-1 receptor domain-containing adapter protein. Chain: a. Fragment: tir domain residues 79-221. Synonym: tir domain-containing adapter protein, adaptor protein wyatt, myd88 adapter-like protein, myd88-2. Engineered: yes. Mutation: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: tirap, mal. Expressed in: escherichia coli. Expression_system_taxid: 511693.

NMR struc:

20 models

Authors:

P.Lavrencic,M.Mobli

Key ref:

M.M.Hughes et al. (2017). Solution structure of the TLR adaptor MAL/TIRAP reveals an intact BB loop and supports MAL Cys91 glutathionylation for signaling. Proc Natl Acad Sci U S A, 114, E6480. PubMed id: 28739909

Date:

27-May-16 Release date: 31-May-17

Protein chain

P58753  (TIRAP_HUMAN) -  Toll/interleukin-1 receptor domain-containing adapter protein from Homo sapiens

Seq: 221 a.a.

Struc: 143 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.?

Proc Natl Acad Sci U S A 114:E6480 (2017)

PubMed id: 28739909

Solution structure of the TLR adaptor MAL/TIRAP reveals an intact BB loop and supports MAL Cys91 glutathionylation for signaling.

M.M.Hughes, P.Lavrencic, R.C.Coll, T.Ve, D.G.Ryan, N.C.Williams, D.Menon, A.Mansell, P.G.Board, M.Mobli, B.Kobe, L.A.J.O'Neill.

ABSTRACT

MyD88 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.