Single molecule localization-based analysis of clathrin-coated pit and caveolar dynamics.

Rui Ma, Martin Štefl, Gerd Ulrich Nienhaus

Nanoscale horizons 2022 Mar 28

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Clathrin-coated pits and caveolae are nanoscale invaginations of the plasma membrane of cells, forming through the assembly of membrane coat and accessory proteins in a tightly regulated process. We have analyzed the development of these membrane coat structures with high spatial and temporal resolution and sensitivity using super-resolution single-molecule localization microscopy (SMLM) on live cells. To this end, we developed a sophisticated clustering and data analysis workflow that automatically extracts the relevant information from SMLM image sequences taken on live cells. We quantified lifetime distributions of clathrin-coated and caveolar structures, and analyzed their growth dynamics. Moreover, we observed hotspots in the plasma membrane where coat structures appear repeatedly. The stunningly similar temporal development of clathrin-coated and caveolar structures suggests that key accessory proteins, some of which are shared between the two types of membrane coat structures, orchestrate the temporal evolution of these complex architectures.

Citation

Rui Ma, Martin Štefl, Gerd Ulrich Nienhaus. Single molecule localization-based analysis of clathrin-coated pit and caveolar dynamics. Nanoscale horizons. 2022 Mar 28;7(4):385-395