Dresden 2014 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 6: Posters: Membranes and vesicles
BP 6.7: Poster
Montag, 31. März 2014, 17:30–19:30, P3
Time-resolved electron tomography reveals how the plasma membrane is reshaped during endocytosis — •Martin Schorb1, Wanda Kukulski1,2, Marko Kaksonen2, and John AG Briggs1,2 — 1Structural and Computational Biology Unit, EMBL, Meyerhofstr. 1, 69117 Heidelberg — 2Cell biology and Biophysics Unit, EMBL, Meyerhofstr. 1, 69117 Heidelberg
Endocytosis is a highly dynamic process that requires a precise temporal and spatial orchestration of multi-component protein modules in order to collect cargo, invaginate the plasma membrane and eventually form an endocytic transport vesicle. Using different pairs of endocytic proteins tagged with GFP and RFP, which act at different stages during endocytosis, we were able to label specific timepoints during the process. By then applying a correlative fluorescence and electron tomography (ET) method, we located specific intermediate stages in 211 individual endocytic budding events, and reconstructed them in 3D.
This dataset provides description of plasma membrane (PM) morphology during the transitions from a plane membrane to tubular invagination, through formation of a constricted neck followed by scission of a vesicle. At each timepoint the presence or absence of key protein players is known. This represents a comprehensive, spatio-temporal description of the plasma membrane topology during endocytosis. A multi-parameter analysis of the membrane profile shapes provides quantitative information about how protein modules of the endocytic machinery coordinate the changes in membrane morphology required for vesicle budding in vivo.