Screen for fission proteins
SMrT templates (see movie) are membrane nanotubes arrayed on a glass coverslip. They can be prepared from diverse lipids and in different sizes, which make them a good mimic of organelles and tubular membrane intermediates generated during vesicle formation in cells. We employ these templates to screen tissue lysates for membrane fission activity, seen as the severing of nanotubes. Using such screens, we expand the repertoire of fission catalysts.
Kamerkar SC, Roy K, Bhattacharyya S, Pucadyil TJ. Biochemistry
Analyze fission mechanisms
Fission requires the enclosing lipid bilayer to be brought to close proximity, which, from theory, represents a distance of separation of 5 nm. Since cellular compartments are of much larger dimensions, fission follows a topological transformation of the limiting membrane into a highly curved tube-like intermediate. Using nanotubes, we monitor dynamics of fission in real time using fluorescence microscopy. Together, these technological developments set the stage for dynamic and correlative analyses of conformational changes in proteins that render them capable of catalyzing membrane fission.
Bassereau P, Jin R, Baumgart T, Deserno M, Dimova R, Frolov VA, Bashkirov PV, Grubmüller H, Jahn R, Risselada HJ, Johannes L, Kozlov MM, Lipowsky R, Pucadyil TJ, Zeno WF, Stachowiak JC, Stamou D, Breuer A, Lauritsen L, Simon C, Sykes C, Voth GA, Weikl TR.
Journal of Physics D Applied Physics
Dar S, Pucadyil TJ. Molecular Biology of the Cell
Dar S, Kamerkar SC, Pucadyil TJ. Nature Cell Biology
Understand fission pathways
Membrane fission appears to be required in a variety of cellular processes - from sorting and trafficking of proteins and lipids in vesicles to organelle biogenesis and division. The hits that emerge from our screen are tested for cellular function by making knock-out models and analyzing morphological changes to various membrane compartments.
Bhattacharyya S, Pucadyil TJ. Protein Science
Deo R, Kushwah MS, Kamerkar SC, Kadam NY, Dar S, Babu K, Srivastava A, Pucadyil TJ. Nature Communications
Kamerkar SC, Kraus F, Sharpe AJ, Pucadyil TJ, Ryan MT. Nature Communications