Electrophoretic Measurements of Protein Folding Intermediates

University Of Texas At Austin, Austin TX

Investigators

Abstract

Professor Jason Shear of the University of Texas Austin will work with Professors Eric Anslyn and Andrew Ellington on a project in bioanalytical chemistry supported by the Analytical and Surface Chemistry Program. The PI initiated a gated ultra-fast electrophoretic separations method in his previous award in the CAREER program. The idea is to cage a molecule, activate it photochemically, do a separation in microseconds and then detect the compounds using light in a time-of-flight analogy. This project centers on the application of that method to the problem of protein folding. Novel photochemical triggering with the help of site-directed mutagenesis to achieve the desired chemistry are being explored. Two model proteins will be used: cytochrome c and apomyoglobin. The project combines analytical and physical chemistry, organic synthesis and molecular biology to develop and evaluate a novel instrument for fast and sensitive detection of protein folding processes. The research is motivated by the question of how protein sequence specifies conformation, a major unanswered question in biology. There is thus a great need for quantitative information about the energetics and dynamics of protein refolding. The role of misfolded proteins is implicated but not well understood in a growing number of diseases such as Alzheimer's disease.