Journal
MOLECULAR BIOLOGY OF THE CELL
Volume 32, Issue 9, Pages 892-902Publisher
AMER SOC CELL BIOLOGY
DOI: 10.1091/mbc.E20-10-0669
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Funding
- National Health, Medical Research Council [APP1102730]
- Swiss National Science Foundation [SNSF 31003A_172969]
- Thurgauische Stiftung fur Wissenschaft und Forschung
- State Secretariat for Education, Research and Innovation
- Deutsche Forschungsgemeinschaft [RO 6238/1-1]
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Endocytic trafficking plays a crucial role in controlling the density of molecules at the plasma membrane, influencing cell interaction with the environment. Understanding how proteins are endocytosed, sorted, and recycled is important for studying cellular processes. The use of photoactivation of fluorescent proteins enables quantitative analysis of endocytosis and intracellular transport.
Endocytic trafficking controls the density of molecules at the plasma membrane and by doing so, the cell surface profile, which in turn determines how cells interact with their environment. A full apprehension of any cellular process necessitates understanding how proteins associated with the plasma membrane are endocytosed, how they are sorted after internalization, and if and how they are recycled to the plasma membrane. To date, it is still difficult to experimentally gain access to this information, even more to do it in a quantitative way. Here we present a toolset based on photoactivation of fluorescent proteins that enabled us to generate quantitative information on endocytosis, incorporation into sorting and recycling endosomes, delivery from endosomes to the plasma membrane, and on the type of vesicles performing intracellular transport. We illustrate these approaches by revealing striking differences in the endocytic trafficking of T-cell receptor and CD4, which bind to the same molecule at the surface of antigen-presenting cells during T-cell activation.
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