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. 

A screen for membrane fission catalysts identifies the ATPase EHD1

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.

The 2018 biomembrane curvature and remodeling roadmap

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

The pleckstrin-homology domain of dynamin is dispensable for membrane constriction and fission

Dar S, Pucadyil TJ. Molecular Biology of the Cell

A high-throughput platform for real-time analysis of membrane fission reactions reveals dynamin function

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. 

Cellular functions and intrinsic attributes of the ATP-binding Eps15 homology domain-containing (EHD) proteins

Bhattacharyya S, Pucadyil TJ. Protein Science

ATP-dependent membrane remodeling links EHD1 functions to endocytic recycling

Deo R, Kushwah MS, Kamerkar SC, Kadam NY, Dar S, Babu K, Srivastava A, Pucadyil TJ. Nature Communications

Dynamin-related protein 1 has membrane constricting and severing abilities sufficient for mitochondrial and peroxisomal fission

Kamerkar SC, Kraus F, Sharpe AJ, Pucadyil TJ, Ryan MT.  Nature Communications