Detection of Polypeptide Interactions Via Periodate Triggered Dopa Crosslinking
| dc.contributor.advisor | Kodadek, Thomas J. | en |
| dc.creator | Burdine, Lyle Jackson | en |
| dc.date.accessioned | 2010-07-12T17:24:15Z | |
| dc.date.available | 2010-07-12T17:24:15Z | |
| dc.date.issued | 2006-08-11 | |
| dc.description.abstract | Protein-Protein interactions mediate most biological function, yet elucidation of the molecular architecture within a cell still remains a formidable challenge for molecular biologist. We have developed a novel, bioorthogonal cross-linking chemistry based upon periodate mediated oxidation of the artificial amino acid 3,4- Dihydroxyphenylalanine to a resultant ortho-quinone. This ortho-quinone was proven capable of capturing either cysteine, lysine, histidine, or a peptidyl alpha -amine in templated chemical reactions. After elucidating the chemistry, we describe utilization of this methodology to map peptide-protein interactions between the 26S proteasome and activation domains as well as the Arp 2/3 complex and the CA peptide. Finally, we present the creation of a chimeric molecule consisting of the biarsencial fluorescent reporter FLAsH conjugated to 3,4-Dihydroxyphenylalanine as a route to deliver 3,4-dihydroxyphenylalanine site specifically to a protein of interest and probe for protein-protein interactions partners in cell lysates. | en |
| dc.format.digitalOrigin | born digital | en |
| dc.format.medium | Electronic | en |
| dc.format.mimetype | application/pdf | en |
| dc.identifier.oclc | 71150591 | |
| dc.identifier.uri | https://hdl.handle.net/2152.5/275 | |
| dc.language.iso | en | en |
| dc.subject | Protein Interaction Mapping | en |
| dc.subject | Intracellular Signaling Peptides and Proteins | en |
| dc.subject | Dihydroxyphenylalanine | en |
| dc.title | Detection of Polypeptide Interactions Via Periodate Triggered Dopa Crosslinking | en |
| dc.type | Thesis | en |
| dc.type.genre | dissertation | en |
| dc.type.material | Text | en |
| thesis.date.available | 2006-08-11 | |
| thesis.degree.department | Graduate School of Biomedical Sciences | en |
| thesis.degree.discipline | Integrative Biology | en |
| thesis.degree.grantor | UT Southwestern Medical Center | en |
| thesis.degree.level | Doctoral | en |
| thesis.degree.name | Doctor of Philosophy | en |