Page 1 |
Save page Remove page | Previous | 1 of 193 | Next |
|
small (250x250 max)
medium (500x500 max)
large ( > 500x500)
Full Resolution
All (PDF)
|
This page
All
Subset |
DEVELOPMENT, SYNTHESIS AND APPLICATIONS OF HIGHLY SENSITIVE
FLUORESCENT PROBES FOR STUDYING PROTEIN TYROSINE
PHOSPHATASES
by
Sayantan Mitra
________________________________________________
A Dissertation Presented to the
FACULTY OF THE GRADUATE SCHOOL
UNIVERSITY OF SOUTHERN CALIFORNIA
In Partial Fulfillment of the
Requirements for the Degree
DOCTOR OF PHILOSOPHY
(CHEMISTRY)
August 2007
Copyright 2007 Sayantan Mitra
Object Description
| Title | Development, synthesis and applications of highly sensitive fluorescent probes for studying protein tyrosine phosphatases |
| Author | Mitra, Sayantan |
| Author email | smitra@usc.edu |
| Degree | Doctor of Philosophy |
| Document type | Dissertation |
| Degree program | Chemistry |
| School | College of Letters, Arts and Sciences |
| Date defended/completed | 2007-06-12 |
| Date submitted | 2007 |
| Restricted until | Restricted until 10 July 2009. |
| Date published | 2009-07-10 |
| Advisor (committee chair) | Barrios, Amy M. |
| Advisor (committee member) |
Bottini, Nunzio Prakash, G.K. Surya |
| Abstract | Protein tyrosine phosphatases (PTPs) constitute an important class of enzymes that are involved in various cellular processes such as mitogenesis, cell growth, signal transduction, immune response and gene transcription. PTP targeted therapeutics have been slow to develop in part, because the substrate selectivity of the PTPs is not well understood and also due to unavailability of suitable probes to study PTP selectivity. Herein is described the design and synthesis of highly fluorescent probes that provide a highly sensitive, continuous assay for PTP activity and are readily incorporated into peptide substrates for high-throughput screening or activity-based assays. These probes are efficiently hydrolyzed by PTPs of both human and bacterial origin. The fluorogenic probe pCAP, was further used to synthesize positional-scanning synthetic combinatorial libraries (PS-SCL) of peptide substrates in order to determine the extended substrate specificities of various PTPs. Such a profile of substrate specificities readily shows both the similarities and differences between PTPs of the same family and those of different families. Furthermore, it was observed that replacing the fluorogenic probe in selective substrates with non hydrolyzable mimics of phosphotyrosine could result in potent, selective inhibitors. The substrate specificities determined from this study would be invaluable in the design of potent, selective PTP substrates and inhibitors for biochemical and therapeutic applications. Furthermore, these probes were used to design peptides for cellular imaging. The viability of the imaging principle using coumarin based probes was checked and various conditions for optimizing internalization of the peptides in cells were analyzed. The imaging technique would allow one to carry out high throughput screening for drugs in cells, and will have a wide impact on the fields of drug development and drug discovery. |
| Keyword | fluorescence; protein tyrosine phosphatases; coumarin; fluorescent amino acids; PTP substrate selectivity; substrate specificity; selective PTP inhibitors; cellular imaging |
| Language | English |
| Part of collection | University of Southern California dissertations and theses |
| Publisher (of the original version) | University of Southern California |
| Place of publication (of the original version) | Los Angeles, California |
| Publisher (of the digital version) | University of Southern California. Libraries |
| Type | texts |
| Legacy record ID | usctheses-m604 |
| Rights | Mitra, Sayantan |
| Repository name | Libraries, University of Southern California |
| Repository address | Los Angeles, California |
| Repository email | http://www.usc.edu/isd/libraries/services/ask_a_librarian/email/ |
| Filename | etd-Mitra-20070710 |
| Archival file | uscthesesreloadpub_Volume35/etd-Mitra-20070710.pdf |
Description
| Title | Page 1 |
| Full text | DEVELOPMENT, SYNTHESIS AND APPLICATIONS OF HIGHLY SENSITIVE FLUORESCENT PROBES FOR STUDYING PROTEIN TYROSINE PHOSPHATASES by Sayantan Mitra ________________________________________________ A Dissertation Presented to the FACULTY OF THE GRADUATE SCHOOL UNIVERSITY OF SOUTHERN CALIFORNIA In Partial Fulfillment of the Requirements for the Degree DOCTOR OF PHILOSOPHY (CHEMISTRY) August 2007 Copyright 2007 Sayantan Mitra |
Comments
Post a Comment for Page 1
