Super-Resolution Imaging of G Protein-Coupled Receptors Using Ground State Depletion Microscopy
| Autor: | Bryan Heit, Stephen S. G. Ferguson, Fabiana A Caetano Crowley |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Cell signaling biology Chemistry G protein Clathrin Cell biology 03 medical and health sciences 030104 developmental biology 0302 clinical medicine Heterotrimeric G protein biology.protein Phosphorylation Receptor Integral membrane protein hormones hormone substitutes and hormone antagonists 030217 neurology & neurosurgery G protein-coupled receptor |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493991204 |
| DOI: | 10.1007/978-1-4939-9121-1_18 |
| Popis: | G protein-coupled receptors (GPCRs) comprise the largest family of integral membrane proteins, which are coupled to heterotrimeric G proteins to influence cell signaling. Subsequent to G protein activation, agonist-stimulated G protein-coupled receptor kinase (GRK) phosphorylation results in the recruitment of β-arrestin proteins, which form both stable and unstable complexes with GPCRs. β-Arrestins when bound to GPCRs not only contribute to the uncoupling of G protein signaling but also to the redistribution of GPCRs to clathrin-coated pits via their association with both clathrin and β2-adaptin facilitating GPCR endocytosis. This allows β-arrestins to couple GPCRs to additional cell signaling proteins allowing a second wave of receptor signaling. Importantly, the β-arrestin-regulated subcellular localization of these complexes also plays a critical role in regulating how these signals are transduced and which proteins are recruited. Here, we describe a methodology for assessing the GPCR subcellular localization by super-resolution microscopy and suggest that this methodology can be extended to the study of GPCR/protein complexes. |
| Databáze: | OpenAIRE |
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