Life Sciences & Biotechnology
Title : | Deep mutational screening for measuring and enhancing protein stability |
Area of research : | Life Sciences & Biotechnology |
Focus area : | Protein Engineering, Computational Biology |
Principal Investigator : | Prof. Raghavan Varadarajan, Indian Institute Of Science, Bangalore, Karnataka |
Timeline Start Year : | 2024 |
Timeline End Year : | 2027 |
Contact info : | varadar@mbu.iisc.ernet.in |
Details
Executive Summary : | While there have been great advances in protein structure prediction, predicting effects of mutations on protein stability remains challenging. We outline a rapid approach to quantitatively estimate relative stabilities of proteins and their mutants without the need of protein purification, and extensions to isolate mutant proteins with improved stability. The protein is expressed on the yeast surface and denatured in the presence of increasing concentrations of denaturant or elevated temperature. The fraction of folded protein is calculated by the ability of the folded or refolded fraction to bind to a conformation specific binding partner, which is monitored by flow cytometry, followed by next generation sequencing of slices of the FACS histogram. Resulting data are analyzed to yield relative stability estimates and identify mutants with enhanced stability. |
Total Budget (INR): | 81,68,600 |
Organizations involved