Event Information
Development of a Protein-Ligand Binding Assay with Multiplex Capabilities
- Abstract:
Currently there is a lack of analytical techniques that can assay direct and indirect protein-ligand interactions on a proteomic scale. Described will be a chemical modification- and mass spectrometry- based approach that has the ability to detect direct and indirect protein-ligand interactions and that can be used in a highly multiplexed fashion for the simultaneous analysis of ligands binding to proteins in complex biological mixtures. A critical part of the described methodology is the detection and quantitation of methionine-containing peptides in the mass spectrometry-based proteomics readout. This work is focused on the development of several experimental strategies to enhance the identification and quantitation of the requisite methionine containing peptides. The initial studies in this work have involved optimization of the LC-MS based proteomics platform using a yeast cell lysate, from which 92 unique peptides from 69 proteins have been detected. The long term goal of this project is to use the described protein-ligand binding assay to identify novel protein targets of the breast cancer drug tamoxifen in order to better understand its mode of action. Preliminary results obtained in an initial application of the methodology to identify the yeast protein targets of tamoxifen have identified two possible new protein targets of the drug. Work is in progress to validate these targets and to further enhance the multiplex capabilities of the technique, which has so far enabled the simultaneous analysis of 69 yeast proteins for binding to tamoxifen.
Preliminary Examination Seminar
Student Exams Seminar