Sushi Madhira

Indian

Biophysical processes at the cellular level are best understood when no ensemble average distorts the overall picture, but rather single molecules are investigated sequentially.

Foerster or Fluorescence resonance energy transfer (FRET) measured between a single donor fluorophore and a single acceptor fluorophore is an ideally suited technique to measure distances and dynamics at single-molecule level. Though single-pair FRET is a powerful tool to study biological molecules undergoing Intra- and Inter-molecular interactions, it is restricted to only one measurable distance per molecule of interest. Owing to this limitation, it cannot reveal intricate details of the global conformational changes during complex biomolecular interactions which are multi-dimensional. For large complexes, using multi-color FRET schemes helps reporting about conformational changes in separate domains of the macromolecule in real time and provides a more comprehensive picture.

In order to expand the single-molecule FRET to study complex molecular interactions, I have assembled a Four-Color Total Internal Reflection Fluorescence (TIRF) Microscope system which allows to measure distances between four fluorophores and thus gives insight into complex molecular dynamics. My research objective is to study DNA-Polymerase interactions and Dynamics of Protein Folding (Chaperones) by employing different multi-color FRET schemes.