Posttranslational Modification of Proteins by ADP-ribosylation Metadata
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- Main Title Posttranslational Modification of Proteins by ADP-ribosylation
Author: Payne, David M. (David Michael)Creator Type: Personal
Chair: Jacobson, MyronContributor Type: PersonalContributor Info: Major Professor
Committee Member: Pirtle, Robert M.Contributor Type: Personal
Committee Member: Harris, Ben G.Contributor Type: Personal
Committee Member: Jacobson, ElaineContributor Type: Personal
Committee Member: Rudick, Michael J.Contributor Type: Personal
Name: North Texas State UniversityPlace of Publication: Denton, TexasAdditional Info: www.unt.edu
- Creation: 1984-12
- Content Description: This work presents the development of a highly sensitive and selective chemical assay for mono(ADP-ribose) residues covalently bound to proteins in vivo. An extensive review of the literature is presented in the introduction of this work. The physiological.functions of mono(ADP-ribosyl)transferase activities associated with certain bacterial toxins (e.g., diphtheria, cholera and pertussis toxins) are well established. However, the roles of endogenous vertebrate transferases are unknown. The elucidation of the roles of these cellular transferases will likely require identification of the physiologically relevant target proteins. Toward this end, it will also be important to identify the types of (ADP-ribose)-protein linkages present in vivo. ADP-ribosylation reactions catalyzed by the different bacterial and vertebrate transferases are specific for different amino acid acceptors in vitro. However, the vertebrate transferases that have been characterized thus far are NAD:arginine mono(ADP-ribosyl)transferases. The work presented here describes the development of a chemical assay for the detection of in vivo modified, ADP-ribosylated proteins containing N-glycosylic linkages to arginine. The assay was applied to the analysis of ADP-ribose residues in adult rat liver. The strategy employed for detection of protein-bound ADP-ribose residues eliminated potential artifacts arising from trapped nucleotides (or their degradation products), since the acid-insoluble material was completely dissolved in a strongly denaturing solution and freed of non-covalently bound nucleotides prior to chemical release from proteins. Thus, the studies presented here demonstrate the unambiguous detection and quantification of protein-bound ADP-ribose residues in adult rat liver. "Arginine-linked" mono(ADP-ribose) residues (31.8 pmol/mg protein) were present in vivo at a level almost 400-fold higher than poly(ADP-ribose). A minor fraction (23%) of the ADP—ribose residues detected were bound via a second more labile linkage with chemical properties very similar to those described previously for carboxlylate esterlinked ADP-ribose. After fractionation of rat liver proteins by gel filtration HPLC, the major peak of "arginine-linked" ADP-ribose residues eluted in the 40-60 kDa region. The later result is consistent with previous suggestions that G-proteins (40-50 kDa) of the adenylate cyclase complex, which are targets for toxin-catalyzed ADP-ribosylation, may also represent target proteins for endogenous transferases.
- Physical Description: xii, 164 leaves: ill.
- Keyword: molecular biology
- Keyword: ribose
- Keyword: amino acids
- Library of Congress Subject Headings: Proteins.
- Library of Congress Subject Headings: Ribose.
Name: UNT Theses and DissertationsCode: UNTETD
Name: UNT LibrariesCode: UNT
- Rights Access: public
- Rights Holder: Payne, David M. (David Michael)
- Rights License: copyright
- Rights Statement: Copyright is held by the author, unless otherwise noted. All rights reserved.
- Thesis or Dissertation
- Accession or Local Control No: 1002779499-Payne
- Call Number: 379 N81d no. 2292
- UNT Catalog No.: b1345109
- OCLC: 14000293
- Archival Resource Key: ark:/67531/metadc331091
- Academic Department: Division of Biochemistry
- Degree Discipline: Molecular Biology
- Degree Level: Doctoral
- Degree Name: Doctor of Philosophy
- Degree Publication Type: disse
- Degree Grantor: North Texas State University