rdf-syntax-grammar
mods:
  schema location: http://www.loc.gov/mods/v3 http://www.loc.gov/standards/mods/v3/mods-3-2.xsd
  title info:
    non sort: A
    title: structural and kinetic study into the role of the quaternary shift in bacillus stearothermophilus phosphofructokinase
  name:
    name part: Mosser, Rockann Elizabeth.
    role:
      role term: creator
  name:
    name part: Reinhart, Gregory D.
    role:
      role term: thesis advisor.
  type of resource: text
  genre: bibliography
  origin info:
    place:
      place term: txu
    date issued: 2011
    issuance: monographic
  language:
    language term: eng
  physical description:
    form: remote
    extent: 1 online resource.
  abstract: Bacillus stearothermophilus phosphofructokinase (BsPFK) is a homotetramer that is allosterically inhibited by phosphoenolpyruvate (PEP), which binds along one dimer-dimer interface. The substrate, fructose-6-phosphate (F6P), binds along the other dimer-dimer interface. The different functional forms BsPFK can take when in the presence of F6P and PEP can be described by the following diproportionation equilibrium: XE + EA <--> XEA + E where XE is the enzyme bound to PEP, EA is the enzyme bound to F6P, E represents the apo enzyme, and XEA is the ternary complex formed when both substrate and inhibitor are bound. Currently in the Protein Data Bank (PDB) there are two relevant forms of wild-type BsPFK, the EA form and the X'E form, which represents the enzyme bound to the PEP analog, phosphoglycolate (PGA). When comparing the EA and the X'E structures, a 7° rotation about the substrate-binding interface is observed and is termed the quaternary shift. The current study uses methyl TROSY NMR to examine the different liganded states of BsPFK, and for the first time structural data for the XEA species is shown. In addition, crystallography was used to obtain the first apo structure of BsPFK. To distinguish between changes associated with the quaternary shift and those associated with the intra-subunit tertiary changes, the variant D12A BsPFK was studied using kinetics, crystallography, and NMR. Crystal structures of apo and PEP bound forms of D12A BsPFK both indicate a shifted structure similar to the X'E form of wild-type. Kinetic studies of D12A BsPFK, when compared to wild-type, show a 50-fold diminished F6P binding affinity, 100-fold enhanced binding affinity, and a similar coupling constant. A conserved hydrogen bond between D12 and T156 takes place across the substrate binding interface in the EA form of BsPFK. The variant T156A BsPFK shows similar binding, coupling, and structural characteristics to D12A BsPFK. PEP still inhibits these variants of BsPFK despite the f…
  note: by Rockann Elizabeth Mosser.
  note: "Major Subject: Biochemistry"
  note: Title from author supplied metadata (automated record created 2011-11-01 09:04:20).
  note: Electronic resource.
  note: Doctor of Philosophy Texas A&M University 2010 http://hdl.handle.net/1969.1/ETD-TAMU-2010-08-8370
  note: Includes bibliographical references.
  note: Text (Dissertation)
  subject:
    topic: Major biochemistry
  subject:
    topic: NMR
  subject:
    topic: Allostery
  subject:
    topic: x-ray crystallography
  subject:
    topic: Energetics
  subject:
    topic: Bacillus stearothermophilus
  subject:
    topic: Enzyme kinetics
  subject:
    topic: PFK
  subject:
    topic: Regulation
  subject:
    topic: Phosphofructokinase
  subject:
    topic: Inhibition
  identifier: http://hdl.handle.net/1969.1/ETD-TAMU-2010-08-8370
  record info:
    record content source: TXA
    record creation date: 111114
    record change date: 20151202144224.0
    record identifier: in00002732974