Protocol for visualizing conditional interaction between transmembrane and cytoplasmic proteins
| Autor: | Masanori Hatakeyama, Takuya Ooki |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Cytoplasm
Science (General) Proximity ligation assay General Biochemistry Genetics and Molecular Biology Cell membrane Q1-390 Cell Line Tumor Protein Biochemistry Protocol medicine Humans Microscopy General Immunology and Microbiology Chemistry Effector General Neuroscience Cell Membrane Membrane Proteins Cell Biology Transmembrane protein Molecular Imaging Cell biology medicine.anatomical_structure Molecular Probes Cell culture Signal transduction Intracellular Protein Binding Signal Transduction |
| Zdroj: | STAR Protocols, Vol 2, Iss 2, Pp 100430-(2021) STAR Protocols |
| ISSN: | 2666-1667 |
| Popis: | Summary This protocol visualizes dynamic interaction between a transmembrane protein and an intracellular protein induced by clusterization/oligomerization of the transmembrane protein. Association-dissociation of the intracellular region of the transmembrane protein with cytoplasmic protein(s) is detected by proximity ligation assay. Since a transmembrane protein often resists extraction, biochemical analysis of its dynamic interaction with cytoplasmic effectors is cumbersome. This protocol quantitatively visualizes protein-protein interaction occurring in the membrane periphery, providing a powerful tool to elucidate signal transduction across the membrane. For complete details on the use and execution of this protocol, please refer to Ooki et al. (2019). Graphical Abstract Highlights • Antibody-mediated cross-linking induces functional clustering of membrane protein • PLA can visualize extracellular-signal-dependent intracellular protein interaction • This protocol is applicable for the analysis of dynamic protein network formation This protocol visualizes dynamic interaction between a transmembrane protein and an intracellular protein induced by clusterization/oligomerization of the transmembrane protein. Association-dissociation of the intracellular region of the transmembrane protein with cytoplasmic protein(s) is detected by proximity ligation assay. Since a transmembrane protein often resists extraction, biochemical analysis of its dynamic interaction with cytoplasmic effectors is cumbersome. This protocol quantitatively visualizes protein-protein interaction occurring in the membrane periphery, providing a powerful tool to elucidate signal transduction across the membrane. |
| Databáze: | OpenAIRE |
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