Single-Molecule Imaging of Signal Transduction via GPI-Anchored Receptors
Lipid rafts have been drawing extensive attention as a signaling platform. To investigate molecular interactions in lipid rafts, we often need to observe molecules in the plasma membranes of living cells because chemical fixation and subsequent immunostaining with divalent or multivalent antibodies may change the location of the target molecules. In this chapter, we describe how to examine dynamics of raft-associated glycosylphosphatidylinositol (GPI)-anchored receptors and interactions of the receptors with downstream signaling molecules by single-particle tracking or single-molecule imaging techniques.
Key wordsSingle-particle tracking Single-molecule imaging GPI-anchored protein Rafts Colocalization Temporal confinement Src family kinase PLCγ IP3 Calcium
This work was supported in part by Grants-in-Aid for Specific Research (B) (No. 24370055) and by Innovative Areas (No. 2311002) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.