Screen for membrane fission catalysts and analyze 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 arrayed membrane nanotubes that mimic such curved intermediates, we screen tissue lysates for fission activity, identify proteins that contribute to such activity, purify them and monitor their dynamics in real time using fluorescence microscopy.
Movie showing fission of membrane nanotubes
Understand physiological functions of membrane fission catalysts
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.
Movie showing vesicle dynamics in cells
Function and regulation of the divisome for mitochondrial fission
Kraus et al. (2021) Nature
Cellular functions and intrinsic attributes of the ATP-binding Eps15 homology domain-containing (EHD) proteins.
Bhattacharyya and Pucadyil (2020) Protein Science
Kamerkar et al. (2019) Biochemistry
Kamerkar and Kraus et al. (2018) Nature Communications
Deo and Kushwah et al. (2018) Nature Communications
Dar and Pucadyil (2017) Molecular Biology of the Cell
Dar et al. (2015) Nature Cell Biology
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