Event Information
Thermodynamic Stability Measurements on Proteins in Complex Mixtures
- Abstract:
The thermodynamic analysis of protein-folding reactions in multi-component protein mixtures is currently a challenge using conventional biophysical techniques. Such analyses are of interest since most such values have been determined for proteins in dilute aqueous solutions, yet cellular conditions are far from pure. The question then arises whether or not proteins have the same thermodynamic parameters in their native environment as they do in a dilute solution. To this end, the viability of performing simultaneous thermodynamic stability determinations of proteins in a cell lysate using a mass spectrometric technique, termed SUPREX (Stability of Unpurified Proteins by Rates of H/D Exchange), was evaluated. SUPREX was then performed on a model mixture of proteins in order to better assess the technical challenges that must be met in order to accomplish the SUPREX analysis of proteins in complex mixtures. Simultaneous analysis of multiple proteins in a model mixture using the SUPREX technique was performed. The results from SUPREX analyses of proteins in the model mixture were compared to values for each protein alone.
A new SUPREX protocol termed Reverse SUPREX was also investigated. Lysozyme was studied using both conventional SUPREX and reverse SUPREX protocols. The thermodynamic parameters obtained from both sets of experiments were compared and appeared to be within the error of these measurements. Reverse SUPREX appears to be a promising technique in the investigation of possible discrepancies in the thermodynamic behavior of proteins in deuterated and protonated solvents.
Master of Science Degree Defense
Student Exams Seminar