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CALM regulates clathrin-coated vesicle size and maturation by directly sensing and driving membrane curvature.

Developmental cell (2015-04-22)
Sharon E Miller, Signe Mathiasen, Nicholas A Bright, Fabienne Pierre, Bernard T Kelly, Nikolay Kladt, Astrid Schauss, Christien J Merrifield, Dimitrios Stamou, Stefan Höning, David J Owen
ABSTRACT

The size of endocytic clathrin-coated vesicles (CCVs) is remarkably uniform, suggesting that it is optimized to achieve the appropriate levels of cargo and lipid internalization. The three most abundant proteins in mammalian endocytic CCVs are clathrin and the two cargo-selecting, clathrin adaptors, CALM and AP2. Here we demonstrate that depletion of CALM causes a substantial increase in the ratio of "open" clathrin-coated pits (CCPs) to "necked"/"closed" CCVs and a doubling of CCP/CCV diameter, whereas AP2 depletion has opposite effects. Depletion of either adaptor, however, significantly inhibits endocytosis of transferrin and epidermal growth factor. The phenotypic effects of CALM depletion can be rescued by re-expression of wild-type CALM, but not with CALM that lacks a functional N-terminal, membrane-inserting, curvature-sensing/driving amphipathic helix, the existence and properties of which are demonstrated. CALM is thus a major factor in controlling CCV size and maturation and hence in determining the rates of endocytic cargo uptake.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-PICALM antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution, Ab1
Sigma-Aldrich
Anti-PICALM antibody produced in rabbit, affinity isolated antibody, buffered aqueous glycerol solution, Ab2

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