APOPTIN AND ITS DERIVATIVES AS MOLECULAR CLUES TOWARDS THE DEVELOPMENT OF NOVEL TYROSINE KINASE INHIBITORS
| dc.contributor.author | Panigrahi, Soumya | |
| dc.contributor.examiningcommittee | Shiu, Robert (Physiology)[Internal Examiner]; Wrogemann, Klaus (Biochemistry and Medical Genetics)[Internal Examiner]; Foley, Ronan (McMaster University, Pathology and Molecular Medicine, Canada)[External Examiner] | en |
| dc.contributor.supervisor | Dodd, Janice (Physiology) [Advisor]; Gibson, Spencer (Biochemistry and Medical Genetics) [Co-advisor] | en |
| dc.date.accessioned | 2009-09-03T14:34:23Z | |
| dc.date.available | 2009-09-03T14:34:23Z | |
| dc.date.issued | 2009-09-03T14:34:23Z | |
| dc.degree.discipline | Physiology | en_US |
| dc.degree.level | Doctor of Philosophy (Ph.D.) | en_US |
| dc.description.abstract | The non-receptor tyrosine kinase activity of fusion gene BCR-ABL derived oncoproteins is the key factor responsible for development and progress of Philadelphia positive (Ph+) chronic myeloid leukemia (CML). In the search for a superior and novel peptide-based inhibitor of Bcr-Abl, here I investigated a naturally occurring molecule, called apoptin. Apoptin is a 13.6 kDa protein derived from chicken anemia virus (CAV) and known to induce apoptosis in a wide range of transformed but not in primary cells. Apoptin is a protein without any reported structural and/or functional homolog and is an interesting candidate to initiate protein-protein interactions and subsequent downstream effects. Initially by an array-based analysis I found that apoptin interacts with the SH3 domain of Abl. By high stringency pull-down and co-immunoprecipitation assays the apoptin and Bcr-Abl interaction was further confirmed. Subsequently, a set of apoptin and Bcr-Abl deletion mutants were used to map this interaction precisely that mainly occurred between a proline rich domain of apoptin and the SH3 domain of Bcr-Abl. I further investigated the role of apoptin on Bcr-Abl. Apoptin was able to modify the phosphorylation of a series of targets (e.g. CrkL, STAT5, c-Myc) downstream of Bcr-Abl kinase. In addition, I used computational algorhythms for protein modeling to study the 3D structure of apoptin and it’s docking with Bcr-Abl at the molecular level. In controlled studies using the 2-pheny-laminopyrimidine derived specific tyrosine kinase inhibitor Imatinib® I found that apoptin has comparable effects on CML cells, suggesting that the interacting segment of the apoptin molecule acts as an adaptor and negatively regulates the Bcr-Abl kinase by deactivating many cell proliferation and anti-apoptotic pathways in CML cells. Briefly, this work provides important insights towards the development of peptide based tyrosine kinase inhibitors as new anti-cancer agents. | en |
| dc.description.note | October 2008 | en |
| dc.format.extent | 4867993 bytes | |
| dc.format.mimetype | application/pdf | |
| dc.identifier.uri | http://hdl.handle.net/1993/3188 | |
| dc.language.iso | eng | en_US |
| dc.rights | open access | en_US |
| dc.subject | Apoptin | en |
| dc.subject | tyrosine-kinase-inhibitor | en |
| dc.title | APOPTIN AND ITS DERIVATIVES AS MOLECULAR CLUES TOWARDS THE DEVELOPMENT OF NOVEL TYROSINE KINASE INHIBITORS | en |
| dc.type | doctoral thesis | en_US |