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Published on: May 8, 2014
Clathrin-coated vesicle formation from isolated plasma membranes
Ishido Miwako1, Sandra L Schmid
1The Scripps Research Institute, Department of Cell Biology, La Jolla, California, USA.
Methods in Enzymology
|January 18, 2006
Summary
Researchers developed a new in vitro assay to study clathrin-coated vesicle (CCV) formation. This assay helps quantify protein requirements for endocytic receptors during CCV assembly.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Clathrin-coated vesicle (CCV) formation is a critical cellular process for endocytosis.
- The precise molecular machinery and event hierarchy governing CCV formation remain incompletely understood.
Purpose of the Study:
- To establish a simplified in vitro system for studying CCV formation.
- To quantitatively assess the protein requirements for various endocytic receptors during CCV assembly.
Main Methods:
- Development of a novel in vitro assay utilizing highly purified rat liver plasma membranes.
- Quantitative evaluation of protein contributions to CCV formation.
Main Results:
- A rapid and straightforward assay for in vitro CCV formation was successfully established.
- The assay allows for precise measurement of protein requirements for distinct endocytic receptors.
Conclusions:
- The developed in vitro assay provides a valuable tool for dissecting the molecular mechanisms of CCV formation.
- This system facilitates the quantitative analysis of protein-receptor interactions in endocytosis.
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