Pucadyil Lab

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

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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

A Screen for Membrane Fission Catalysts Identifies the ATPase EHD1

Kamerkar et al. (2019) Biochemistry 

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

Kamerkar and Kraus et al. (2018) Nature Communications

ATP-dependent membrane remodeling links EHD1 functions to endocytic recycling

Deo and Kushwah et al. (2018) Nature Communications 

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

Dar and Pucadyil (2017) Molecular Biology of the Cell

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

Dar et al. (2015) Nature Cell Biology 

Click here to view Pucadyil Lab publications on Pubmed