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  schema location: http://www.loc.gov/mods/v3 http://www.loc.gov/standards/mods/v3/mods-3-2.xsd
  title info:
    title: Mutational effects on the conformational stability and activity of Ribonuclease T1
  name:
    name part: Shirley, Bret Allen
    name part: 1962-
    role:
      role term: creator
  name:
    name part: Pace, C. Nick
    role:
      role term: degree supervisor.
  name:
    name part: Kelly, Jeffery W.
    role:
      role term: degree committee member.
  name:
    name part: Peterson, David O.
    role:
      role term: degree committee member.
  name:
    name part: Pettigrew, Donald W.
    role:
      role term: degree committee member.
  name:
    name part: Texas A & M University
    role:
      role term: degree granting institution.
  type of resource: text
  genre: bibliography
  genre: Academic theses
  origin info:
    place:
      place term: xx
    date issued: 1990
    issuance: monographic
  language:
    language term: eng
  physical description:
    form: print
    extent: x, 99 leaves : illustrations ; 29 cm
  abstract: Ribonuclease T1 (RNase T1) and fourteen mutants were prepared from a chemically synthesized gene, cloned and expressed in Escherichia coli. The wild-type RNase T1 prepared from E. coli was identical in every functional and physical property examined to RNase T1 prepared from Aspergillus oryzae. Urea and thermal unfolding experiments show that the amino acid substitution Gln25 [to] Lys is 0.9 kcal/mol more stable and Glu58 [to] Ala is 0.8 kcal/mol less stable than wild-type RNase T1. In the mutant Glu58 [to] Ala (K25) (i.e. the double mutant), these contributions cancel and the stability does not differ significantly from that of wild-type RNase T1. The activity of Gln25 [to] Lys is identical with that of wild-type RNase T1. However, the activities of Glu58 [to] Ala and Glu58 [to] Ala (K25) are 7% of wild-type when GpC hydrolysis is measured (due to a 35-fold decrease in k[cat]), and 37% of wild-type when RNA hydrolysis is measured. Thus, Glu58 is important, but not essential to the activity of RNase T1. Urea and thermal unfolding curves were determined for eleven mutants that remove specific hydrogen bonds from the native state structure of RNase T1. These studies show that the deletion of a partner from a neutral-neutral hydrogen bond pair reduces stability by 0.6 to 1.5 kcal/mol, and the deletion of a partner from a neutral-charged pair reduces stability by as much as 2 kcal/mol. The most surprising finding was that for two of these mutants, the removal of a hydrogen bonding partner resulted in an increase in the conformational stability of RNase T1. These results suggest that hydrogen bonds make a significant contribution to the conformational stability of globular proteins.
  note: Typescript (photocopy).
  note: Vita.
  note: "Major subject: Biochemistry."
  note: Ph. D. Texas A & M University 1990
  note: Includes bibliographical references.
  subject:
    topic: Nucleoproteins
    topic: Analysis
  subject:
    topic: Recombinant proteins
  subject:
    topic: Hydrogen bonding
  subject:
    topic: Major biochemistry
  identifier: http://gateway.proquest.com/openurl
  identifier: http://proxy.library.tamu.edu/login
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    location:
      url: http://proxy.library.tamu.edu/login
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    location:
      url: http://gateway.proquest.com/openurl
  record info:
    record content source: TXA
    record creation date: 910905
    record change date: 20200904152533.0
    record identifier: in00001043022
    language of cataloging:
      language term: eng