With this award, the Chemical Measurement and Imaging Program in the Division of Chemistry is supporting Professors Tao and Zhu at Purdue University to develop and implement a platform for identification and quantification of phosphoproteomes of interest. Reversible protein phosphorylation regulates a number of biological processes, such as onset of many diseases and plant signaling. While it has been found extensive, at any given time only a small percentage of phosphorylated proteins may change their phosphorylation status in response to a particular cellular event, which makes it extremely challenging for detection and quantitative measurement of phosphorylated proteins of relevant. The proposed proteomic platform aims at a versatile and cost-effective analytical tool. The technique will be applied to identify important phosphoproteins involved in osmosis, water, temperature, cold, and nutrient deficiency stress in Arabidopsis. In addition to providing interdisciplinary learning experience to both graduate and undergraduate students, the PIs also plan to promote teaching, training, learning, and discovery through the development of a new summer undergraduate lab course to engage students in a genuine research project that emphasizes the utilization of modern analytical methods and to enhance students' academic opportunities.
The project will develop a novel proteomic platform that addresses two major analytical questions related to large-scale protein phosphorylation analyses: how to quantitatively visualize and detect phosphorylated proteins in a complex sample and how to identify phosphoproteins of interest and their sites of phosphorylation. The core of the platform is novel chemical reagents that are multi-functionalized for high specificity and selectivity toward phosphoproteins in complex biological samples such as whole cell extracts. The platform will allow for the selection of phosphoproteins of interest that are further subjected to tandem mass spectrometry to get sequencing information and sites of phosphorylation. The technique seeks to provide researchers with a useful technique for routine phosphorylation analyses. Major applications include quantitative measurement of phosphorylation, assays of kinase and phosphatase activity, screening for kinase/phosphatase inhibition, and detection of in vivo phosphorylation in different cellular states.
