Georgetown University LogoGeorgetown University Library LogoDigitalGeorgetown Home
    • Login
    View Item 
    •   DigitalGeorgetown Home
    • Georgetown University Institutional Repository
    • Georgetown College
    • Department of Chemistry
    • Graduate Theses and Dissertations - Chemistry
    • View Item
    •   DigitalGeorgetown Home
    • Georgetown University Institutional Repository
    • Georgetown College
    • Department of Chemistry
    • Graduate Theses and Dissertations - Chemistry
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Interactions of quinolinols, peptides and SNAP-25 with the protease domain of botulinum neurotoxin A

    Cover for Interactions of quinolinols, peptides and SNAP-25 with the protease domain of
      botulinum neurotoxin A
    View/Open
    View/Open: laiHuiguo.pdf (2.2MB) Bookview

    Creator
    Lai, Huiguo.
    Description
    Thesis (Ph.D.)--Georgetown University, 2010.; Includes bibliographical references.; Text (Electronic thesis) in PDF format. SNAP-25, an intrinsically disordered protein, assembles with syntaxin and synaptobrevin to form a SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) complex, a four-helix bundle that plays a key role in the exocytosis of synaptic vesicles. Botulinum neurotoxins specifically cleave one of the SNARE proteins, resulting in the inhibition of membrane fusion. Small molecule inhibitors are being evaluated as potential therapeutic counter-measurement to the toxin's deleterious action on the neuromuscular junctions.; In order to understand the conformational changes exhibited by SNAP-25 prior to and during SNARE complex formation, and its biological function, we developed a method of investigating the conformational changes of SNAP-25 in the absence and presence of Zn2+-depleted BoNT/A LC (botulinum neurotoxin A light chain), using FRET (fluorescence resonance energy transfer) between EGFP (enhanced green fluorescent protein) and CCPGCC-ReAsH fused to SNAP-25 at specific locations. The apparent distances between the C-terminus and the N-terminus, the C-terminus and the D140, and the N-terminus and the D140 of the SNAP-25 protein were determined based on the FRET efficiency in the absence and presence of Zn2+-depleted BoNT/A LC. Various mutants of SNAP-25(A195S) were genetically engineered and used to measure the tertiary structures of SNAP-25(A195S) in the absence and presence of active BoNT/A LC. Shorter apparent distances in the presence of BoNT/A LC suggest that the SNAP-25 folds around BoNT/A LC in the binary complex. FRET is a useful tool to investigate the conformational changes of other intrinsically disordered proteins.
    Permanent Link
    http://hdl.handle.net/10822/552881
    Date Published
    2010
    Subject
    Biochemistry
    Type
    thesis
    Publisher
    Georgetown University
    Collections
    • Graduate Theses and Dissertations - Chemistry
    Metadata
    Show full item record

    Related items

    Showing items related by title, author, creator and subject.

    • Cover for Mechanisms of SNAP-25 Association with Botulinum Neurotoxin Light Chain A and SNARE Proteins

      Mechanisms of SNAP-25 Association with Botulinum Neurotoxin Light Chain A and SNARE Proteins 

      Sultani, Mashal (Georgetown University, 2012)
      Botulism neurotoxins (BoNTs), a family of neurotoxins produced by the anaerobic bacterium Clostridium botulinum, are the most poisonous biological toxins known to man and are listed as category A bioterrorism agents by the ...
    Related Items in Google Scholar

    Georgetown University Seal
    ©2009 - 2023 Georgetown University Library
    37th & O Streets NW
    Washington DC 20057-1174
    202.687.7385
    digitalscholarship@georgetown.edu
    Accessibility
     

     

    Browse

    All of DigitalGeorgetownCommunities & CollectionsCreatorsTitlesBy Creation DateThis CollectionCreatorsTitlesBy Creation Date

    My Account

    Login

    Statistics

    View Usage Statistics

    Georgetown University Seal
    ©2009 - 2023 Georgetown University Library
    37th & O Streets NW
    Washington DC 20057-1174
    202.687.7385
    digitalscholarship@georgetown.edu
    Accessibility