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  • Molecular Mimicry in Innate Immunity:: Structure of A Bacterial TIR Virulence Factor

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    This document summarizes research on molecular mimicry in innate immunity. It discusses the structure of bacterial TIR virulence factors and their similarity to eukaryotic TIR domains. It describes how bacterial TIR-like proteins can interact with host signaling proteins like MyD88, interfering with toll-like receptor signaling pathways and inhibiting the host immune response. The document also presents structural and functional studies of prokaryotic TIR-like proteins from various bacteria and discusses their role in virulence. Finally, it proposes that bacterial TIR mimics could be targeted for drug development to inhibit pathogen interference with the host immune system.

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    This document summarizes research on molecular mimicry in innate immunity. It discusses the structure of bacterial TIR virulence factors and their similarity to eukaryotic TIR domains. It describes how bacterial TIR-like proteins can interact with host signaling proteins like MyD88, interfering with toll-like receptor signaling pathways and inhibiting the host immune response. The document also presents structural and functional studies of prokaryotic TIR-like proteins from various bacteria and discusses their role in virulence. Finally, it proposes that bacterial TIR mimics could be targeted for drug development to inhibit pathogen interference with the host immune system.

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    52 views20 pages

    Molecular Mimicry in Innate Immunity:: Structure of A Bacterial TIR Virulence Factor

    Uploaded by

    jamie pascal
    This document summarizes research on molecular mimicry in innate immunity. It discusses the structure of bacterial TIR virulence factors and their similarity to eukaryotic TIR domains. It describes how bacterial TIR-like proteins can interact with host signaling proteins like MyD88, interfering with toll-like receptor signaling pathways and inhibiting the host immune response. The document also presents structural and functional studies of prokaryotic TIR-like proteins from various bacteria and discusses their role in virulence. Finally, it proposes that bacterial TIR mimics could be targeted for drug development to inhibit pathogen interference with the host immune system.

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    Molecular Mimicry in Innate Immunity:

    Structure of a Bacterial TIR


    Virulence Factor

    Jaime Pascual, Burnham Institute


    Toll-like receptor signaling pathway

    κ
    Suzuki, Trends in Immunology (2002)
    Toll-like receptor activation mechanism
    DeGrado, Structure (2008)
    Crystal structure of monomeric human
    Toll-like receptor 1 TIR domain

    TIR 3D fold belongs


    PDB_ID: 1FYV
    to the flavodoxin-like
    α/β family
    that includes
    bacterial proteins
    Model for TIR-TIR signaling interaction:
    homotypic via E-helix and heterotypic via BB loop

    Jiang, PNAS (2006)


    Striking a balance through host mimicry:
    Myxoma virus M11L virulence factor:
    a Bcl-2 mimic binding and antagonizing pro-apoptotic Bax & Bak

    Colman, Mol Cell (2007)


    Sequence alignment between prokaryotic & eukaryotic TIRs
    Native protein crystal of
    P. denitrificans TLP TIR domain
    Crystallization conditions at 4° C: Dataset collected in-house:
    Salt: 0.2 M Ammonium sulfate Resolution: 2.50 Å
    Buffer: 0.1 M Sodium cacodylate pH 6.0 Space group: P 212121
    Precipitant: 31% (w/v) PEG 8000 Unit cell: a=80.8; b=85.6; c=89.6
    PdTLP TIR structure has a TIR fold
    Superposition of PdTLP-TIR domain onto human TLR1-TIR domain
    (Cα RMSD = 2.8 Å)
    PdTLP TIR crystal dimer:
    Exposed BB loops available for heterotypic interactions

    Chain B
    DD loop

    BB loop
    EE loop

    Chain A

    BB loop
    Gel filtration chromatograms for PdTLP
    aa1 Leu145 aa299
    Helical bundle TIR
    pI 10.2 Full length pI 9.6 pI 5.7

    600

    158kDa 44kDa 17kDa


    500
    Dimer
    400

    Full length
    mAU

    300

    200
    (33 kDa)
    100

    0
    0 20 40 60 80 100
    ml

    -10

    -20 Oligomer
    -30 N-terminal
    mAU

    Helical bundle
    -40

    -50

    -60

    -70
    (16 kDa)
    -80
    0 20 40 60 80 100
    ml
    600

    500
    Monomer
    400
    C-terminal
    mAU

    300

    200 TIR
    100
    (17 kDa)
    0

    -100
    0 20 40 60 80 100
    ml
    Dimerization Interface Determined Using
    Deuterium Exchange Mass Spectrometry (DXMS)

    PdTIR -
    monomer

    H H
    B B full length
    PdTLP -
    dimer
    DXMS of PdTIR vs PdTLP:
    Same protected interface in PdTLP as in the PdTIR crystal dimer
    DXMS of PdTIR vs PdTLP:
    Same protected interface in PdTLP as in the PdTIR crystal dimer
    Co-immunoprecipitation between
    MyD88 and PdTLP TIR domains

    ds
    88

    a
    ac
    t

    be
    yD
    vc

    Sm

    ph
    M
    p
    M.W.

    Em

    Se
    4µg 2µg 4µg 2µg
    (kDa)

    *
    37
    IP-Myc
    MyD88
    IB-HA
    26
    *
    *
    19
    IP-Myc
    IB-Myc
    PdTLP 15

    37 *
    MyD88 Lysate
    IB-HA
    26
    Smac
    Functional studies on bacteria TLP proteins

    Salmonella enterica (Newman, et al., 2006) :


    delayed disease effects of a TIR-like knockout strain infecting mice

    Brucella abortus (Salcedo, et al., 2008) :


    a TIR-like knockout strain infecting human dendritic cells produced increased
    levels of pro-inflammatory cytokine secretion

    Escherichia coli (Cirl, et al., 2008) :


    TIR-like virulence factor directly interacts with human MyD88
    Drugability of the TLR pathway

    Beutler, Nature (2004)


    BB loop mimic as inhibitor of prot-prot inter:
    a low molecular weight pyrrolidine interacts with MyD88 TIR

    Rebek & Bartfai, PNAS (2006)


    Model for virulence action by TIR mimicry
    Healthy macrophage Infected macrophage
    PAMP PAMP

    TLR
    LRR

    TLR

    TIR
    Death Domain

    Helical bundle
    MyD88

    TIR virulence factor

    kinase
    IRAK
    TIR

    MyD88

    NF-кB activation NF-кB activation


    Acknowledgements
    Group

    Siew Leong Chan & Lieh Yoon Low


    Collaborators

    Eugenio Santelli (Robert C.


    Liddington)

    Gaelle Le Negrate (John C. Reed)

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