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Mechanisms of clathrin-mediated endocytosis

Key Points

  • Clathrin-coated endocytic vesicles are produced by a complex modular protein machinery that transiently assembles on the plasma membrane. This machinery selects and concentrates cargo molecules and shapes the membrane into a vesicle.

  • Forces arising within the membrane during deformation counteract forces generated by the endocytic protein modules. Physical parameters such as membrane tension and rigidity control the dynamics of clathrin-mediated endocytosis.

  • Many endocytic proteins bind phosphoinositides, which are critical for organizing the sequence of protein assembly throughout endocytosis.

  • The endocytic machinery is evolutionarily ancient and highly conserved, but it has adapted to varying force requirements in different lineages.

Abstract

Clathrin-mediated endocytosis is a key process in vesicular trafficking that transports a wide range of cargo molecules from the cell surface to the interior. Clathrin-mediated endocytosis was first described over 5 decades ago. Since its discovery, over 50 proteins have been shown to be part of the molecular machinery that generates the clathrin-coated endocytic vesicles. These proteins and the different steps of the endocytic process that they mediate have been studied in detail. However, we still lack a good understanding of how all these different components work together in a highly coordinated manner to drive vesicle formation. Nevertheless, studies in recent years have provided several important insights into how endocytic vesicles are built, starting from initiation, cargo loading and the mechanisms governing membrane bending to membrane scission and the release of the vesicle into the cytoplasm.

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Figure 1: Stages of clathrin-mediated endocytosis and the associated modular machinery.
Figure 2: Initiation of membrane budding during endocytosis is mediated by the pioneer module.
Figure 3: Mechanism of clathrin coat-mediated and actin-mediated membrane bending.
Figure 4: Membrane fission by dynamin and BAR proteins followed by vesicle uncoating.

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Acknowledgements

The authors dedicate this Review to the memory of C. Merrifield, who prematurely passed away on 29 November 2017. In many aspects, C. Merrifield's contribution to the understanding of clathrin-mediated endocytosis has been essential. He was, through his genuine scientific interest, kindness and passion, a true gentleman. The authors will remember him vividly. A.R. acknowledges funding from a Human Frontier Science Program Young Investigator Grant (RGY0076-2008), a European Research Council starting (consolidator) grant (311536-MEMFIS) and the Swiss National Science Foundation (grants 131003A_130520 and 131003A_149975). M.K. acknowledges funding from the Swiss National Science Foundation (grant 31003A_163267).

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Both authors contributed equally to all aspects of the article (researching data for the article and substantial contribution to the discussion of content, writing, review and editing of the manuscript before submission).

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Glossary

Turgor pressure

Osmotic pressure within plant and fungal cells confined within a cell wall.

BAR domain

A crescent-shaped dimeric protein domain that binds membranes with its curved surface and thereby either senses membrane curvature or bends the membrane.

Membrane tension

In-plane force counteracting membrane surface expansion.

Elastic energy

Energy required to deform an elastic material. For lipid membranes, it contains a term for bending and a term for stretching, both taking the form of the energy associated with a harmonic spring: a constant called the modulus or rigidity, multiplied by the shape change to the square. Thus, bending energy is the bending rigidity multiplied by membrane curvature to the square, whereas stretching energy is the compressibility modulus multiplied by the area difference to the square. The elastic energy of the membrane is the sum of these two terms.

Amphipathic helix

A short polypeptide, typically between 10 and 20 amino acids in length, that contains hydrophobic and hydrophilic residues. This polypeptide spontaneously folds into an α-helix when binding to a lipid membrane. In this configuration, all hydrophobic residues are aligned on the cylindrical face of the helix that is buried in the bilayer whereas the hydrophilic moieties are aligned on the hydrated face.

Fluorescence recovery after photobleaching

(FRAP). Microscopy method for measuring local exchange of fluorescently labelled molecules.

Type I myosins

A highly conserved subfamily of monomeric myosin motors involved in cell motility and membrane traffic.

Entropic forces

Forces that arise while the system tries to maximize its entropy. These forces typically arise from frustrated thermal fluctuations, which will then counteract the constraints by applying forces onto them. The pressure of an ideal gas is an entropic force. In lipid membranes, repulsive forces between closely apposed bilayers (less than a few tens of nanometres) — known as Helfrich forces — are entropic forces. They arise from thermal undulations of the bilayer surface. In polymer physics, thermal fluctuations usually lead to the folding of the polymer molecule into globular conformations. If one pulls on both ends of the molecule, an entropic force is felt.

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Kaksonen, M., Roux, A. Mechanisms of clathrin-mediated endocytosis. Nat Rev Mol Cell Biol 19, 313–326 (2018). https://doi.org/10.1038/nrm.2017.132

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