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  • Harrison Powerpoint Pt2

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    bioe07011
    51 tayangan33 halaman
    This document summarizes the process of membrane fusion used by enveloped viruses like influenza to infect cells. It begins with an overview of the viral and cellular membranes fusing together. It then examines the sequence of events in viral membrane fusion, from the prefusion to postfusion stages. Key proteins involved in fusion, like the class I viral fusion proteins of influenza, HIV, Ebola, are introduced. The document focuses on influenza virus, describing the HA and NA proteins and HA's role in receptor binding, antigenic variation, and membrane fusion. It outlines HA's cleavage and low pH-triggered conformational changes that expose the fusion peptide and drive membrane fusion. Single-virion fusion experiments are described that probe the kinetics and

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    This document summarizes the process of membrane fusion used by enveloped viruses like influenza to infect cells. It begins with an overview of the viral and cellular membranes fusing together. It then examines the sequence of events in viral membrane fusion, from the prefusion to postfusion stages. Key proteins involved in fusion, like the class I viral fusion proteins of influenza, HIV, Ebola, are introduced. The document focuses on influenza virus, describing the HA and NA proteins and HA's role in receptor binding, antigenic variation, and membrane fusion. It outlines HA's cleavage and low pH-triggered conformational changes that expose the fusion peptide and drive membrane fusion. Single-virion fusion experiments are described that probe the kinetics and

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    Attribution Non-Commercial (BY-NC)
    Unduh sebagai PDF, TXT atau baca online dari Scribd
    Tandai sebagai konten tidak pantas
    51 tayangan33 halaman

    Harrison Powerpoint Pt2

    Diunggah oleh

    bioe07011
    This document summarizes the process of membrane fusion used by enveloped viruses like influenza to infect cells. It begins with an overview of the viral and cellular membranes fusing together. It then examines the sequence of events in viral membrane fusion, from the prefusion to postfusion stages. Key proteins involved in fusion, like the class I viral fusion proteins of influenza, HIV, Ebola, are introduced. The document focuses on influenza virus, describing the HA and NA proteins and HA's role in receptor binding, antigenic variation, and membrane fusion. It outlines HA's cleavage and low pH-triggered conformational changes that expose the fusion peptide and drive membrane fusion. Single-virion fusion experiments are described that probe the kinetics and

    Hak Cipta:

    Attribution Non-Commercial (BY-NC)
    Unduh sebagai PDF, TXT atau baca online dari Scribd
    Tandai sebagai konten tidak pantas
    dari 33

    Enveloped viruses acquire membrane ! by budding!

    HA NA

    RNP

    alphaviruses (e.g., Sinbdis virus)

    influenza virus

    alphaviruses

    influenza virus

    Enveloped viruses penetrate cells by fusion of viral and cellular membranes Influenza: enters through endosomes, depends on low pH of endosome to initiate fusion process HIV: can fuse at cell surface, depends on receptor (CD4) and co-receptor to initiate fusion process

    What is membrane fusion?


    two bilayers hemifusion fusion pore

    Sequence of events in viral membrane fusion


    Cell

    Virus

    Prefusion

    Extended intermediate

    Collapse of intermediate

    Hemifusion

    Fusion pore (postfusion)

    "Class 1" viral fusion proteins


    Orthomyxoviridae Influenza A Retroviridae HIV-1 Filoviridae Ebola

    Envelope glycoprotein
    Receptor binding domain Fusion molecule

    TM

    Inuenza virus particle!

    Electron micrograph! of inuenza virus

    hemagglutinin (HA) neuraminidase (NA)!


    (X 5 million = golf tee)!

    (X 1 million = golf ball)!

    500 !

    hemagglutinin (HA): ! three functions!


    1. receptor binding! 2. antigenic variation! 3. fusion!

    hemagglutinin (HA): ! synthesized as precursor:!

    Influenza virus hemagglutinin

    viral membrane

    Influenza virus hemagglutinin

    Influenza virus hemagglutinin


    sialic-acid binding site

    -C HA1 HA2 N-

    HA2 C-

    -N HA1

    Low-pH triggered conformational change


    TM fp

    HA2 Nfusion peptide

    HA2 TM <

    - HA1 N

    HA trimer: pH 7

    pH<5.5

    HA monomer: pH 7 pH<5.5

    HA undergoes two irreversible changes ...

    energy

    HA0

    post-translational cleavage

    neutral pH conformation metastable low pH conformation

    ...the second change catalyzes membrane fusion


    cell

    virus

    Fusion mechanism A. Cleave precursor (prime) B. Localize virus to cell (by receptor binding) C. Trigger refolding (by co-receptor, low pH, etc.) 1. Expose fusion peptide 2. Insert fusion peptide into target membrane 3. Fold back to bring together target and viral membranes

    ...the second change catalyzes membrane fusion


    cell

    virus

    trigger

    expose

    insert

    fold back

    cap

    Similarity among post-fusion structures


    TM anchors and fusion peptides

    low pH influenza eHA2

    SIV gp41 (NMR)

    ebola GP2

    HTLV-1 gp21

    Gal McGill molecularmovies.com

    Single-virion fusion measurements

    ganglioside

    fluorescein pH sensor

    100 nm

    Floyd et al, 2008

    Histograms of times to hemifusion (lipid mixing) and pore formation (content mixing)

    Two-parameter kinetic scheme: N, k


    N sequential steps

    A X1 X2 XN-1 H
    OR N independent parallel steps

    AH AH AH ....
    k k

    Best fits to observed histograms


    N=3 (time to hemifusion) interpret as number of HA trimers required to create a fusion pore

    N=1 (exponential) (hemifusion decay: time from hemifusion to pore formation for individual particles -- single rate-limiting step)

    pH dependence

    pH dependence k

    Influenza HA fusion

    H+

    Native

    H+

    HA1 open

    HA2 extended

    HA2 collapse ~20 sec

    Mutations here accelerate fusion

    Influenza HA fusion

    H+

    Native

    H+

    HA1 open

    HA2 extended

    HA2 collapse ~20 sec

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