Collaborative Research: Single Cell Proteomics

Louisiana State University, Baton Rouge LA

Investigators

Abstract

This award provides support for a joint project aimed at the development of aggressive new analytical strategies for protein sequence analysis based on mass spectrometry. The effort involves investigators at Louisiana State University, who are accomplished in the manufacture of micrometer-sized devices, and their collaborator at the University of Cincinnati, who is skilled in high resolution mass spectrometry. Microfabrication techniques will be used to develop integrated micro-systems expected to permit analysis of the proteins in a single cell. The use of microfludics will help maintain the proteins at a high enough concentration to permit analysis even though the amount of each protein found in a cell is on the order of 3000 molecules. The devices to be used in this project are built using polymers and micromanufacturing techniques to fabricate high-aspect-ratio molding dies for hot embossing polymers. The devices will be tested and refined through studies of chloroplast-derived membranes thought to contain at least 150 different proteins. Within the context of the proposed effort, these studies will be extended to examine the proteome, i.e., the entire complement of proteins, of intact chloroplasts of Arabadopsis and of entire cells of the blue-green algae Synechocystis. If fully successful, the instrumentation will allow biochemical analyses of individual cells and thus contribute to a better understanding of the relationship of gene expression and cellular phenotype.