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:
    title: Solubilization and functional analysis of the lambda holin
  name:
    name part: Deaton, John Franklin
    name part: 1974-
    role:
      role term: creator
  name:
    name part: Young, Ryland F.
    role:
      role term: thesis advisor.
  type of resource: text
  genre: bibliography
  origin info:
    place:
      place term: txu
    date issued: 2004
    issuance: monographic
  language:
    language term: eng
  physical description:
    form: remote
    extent: 1 online resource.
  abstract: The 105aa lambda S protein is the prototype holin, S accumulates in the cytoplasmic membrane during late gene expression until, at a time programmed into its primary structure, it disrupts the membrane and allows the lambda lysozyme, R, to attack the cell wall. In this study, a zwitterionic detergent Empigen BB, was used to extract and purify the lambda holin S. In Empigen BB, CD analysis on S gave 54% alpha helical content, consistent with 3 TM domains, which has been reported by other in vivo studies. Empigen BB-purified S can be exchanged into a chaotropic solution by dialysis and reconstituted into preformed lipid vesicles for activity assays. When diluted to fluorescein-loaded suspensions of liposomes, different chaotrope-solubilized S alleles caused dye release reflective of their in vivo phenotypes. The problem was the low efficiency of delivery of S to the liposomes. Unfortunately, dye loaded liposomes are highly sensitive to any detergent, making it necessary to find other ways to solubilize S. GroEL, a chaperonin from E. coli, is responsible for folding and refolding globular proteins in vitro. It has also been reported that GroEL improves the ability of a membrane protein synthesized in vitro to insert post-translationally into liposomes. This work will investigate the behavior of GroEL towards membrane proteins. The first of two membrane proteins studied in this respect is Bacteriorhodopsin (BR), a membrane proton pump, from H.halibium. The second is the105aa S protein, a prototype holin from bacteriophage lambda. Holin and BR subjected to detergent removal in the presence of GroEL remained in solution, while in the control sample (without GroEL) S and BR precipitated. "GroELsolubilized" holin still retained its lesion forming activity and solubilized BR maintained its proton pumping ability, detected by using a liposome dye activity assay unique to each protein. This approach may be applicable to other systems requiring detergent- or chaotrope-free prep…
  note: by John Franklin Deaton.
  note: Vita.
  note: Abstract.
  note: "Major Subject: Biochemistry"
  note: Title from author supplied metadata (automated record created on Oct. 15, 2004.)
  note: Electronic resource.
  note: System requirements: Adobe Acrobat Reader.
  note: Ph. D. Texas A & M University 2004.
  note: Includes bibliographical references.
  note: Text (Dissertation).
  subject:
    topic: Major biochemistry
  subject:
    topic: GroEL
  subject:
    topic: chaperonin
  subject:
    topic: membrane protein
  subject:
    topic: holin
  identifier: http://hdl.handle.net/1969.1/1211
  record info:
    record content source: TXA
    record creation date: 050303
    record change date: 20151101052852.0
    record identifier: in00002038189