title: Single-Molecule Imaging of Signal Transduction via GPI-Anchored Receptors.
abstract: 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.
authors.affiliation: Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Institute for Frontier Medical Sciences, Kyoto University, Kyoto, 606-8507, Japan. ksuzuki@frontier.kyoto-u.ac.jp.|National Centre for Biological Sciences (NCBS)/Institute for Stem Cell Biology and Regenerative Medicine (inStem), Bangalore, 560065, India. ksuzuki@frontier.kyoto-u.ac.jp.
authors.forename: Kenichi G N
authors.initials: KG
authors.lastname: Suzuki
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000494:pharmacology; D018957:CD59 Antigens / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D005456:Fluorescent Dyes; D058851:GPI-Linked Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D053768:Metal Nanoparticles; D057054:Molecular Imaging / Q000379:methods*; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D015398:Signal Transduction; D013194:Staining and Labeling
chemical_list: D000911:Antibodies, Monoclonal; D018957:CD59 Antigens; D005456:Fluorescent Dyes; D058851:GPI-Linked Proteins; D011956:Receptors, Cell Surface; D015544:Inositol 1,4,5-Trisphosphate; D002118:Calcium
keywords: Calcium; Colocalization; GPI-anchored protein; IP3; PLCγ; Rafts; Single-molecule imaging; Single-particle track ing; Src family kinase; Temporal confinement