Single Cell GFP-Trap Reveals Stoichiometry and Dynamics of Cytosolic Protein Complexes
| Autor: | Jacob Piehler, Tim Wedeking, Christian Richter, Changjiang You, Maniraj Bhagawati, Sara Löchte |
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| Rok vydání: | 2015 |
| Předmět: |
In situ
Materials science Lysis Total internal reflection fluorescence microscope Surface Properties Mechanical Engineering Analytical chemistry Bioengineering General Chemistry Condensed Matter Physics Single Molecule Imaging Protein–protein interaction Green fluorescent protein Cytosol Microscopy Fluorescence Multiprotein Complexes Microscopy Biophysics Humans General Materials Science Single-Cell Analysis |
| Zdroj: | Nano Letters. 15:3610-3615 |
| ISSN: | 1530-6992 1530-6984 |
| DOI: | 10.1021/acs.nanolett.5b01153 |
| Popis: | We developed in situ single cell pull-down (SiCPull) of GFP-tagged protein complexes based on micropatterned functionalized surface architectures. Cells cultured on these supports are lysed by mild detergents and protein complexes captured to the surface are probed in situ by total internal reflection fluorescence microscopy. Using SiCPull, we quantitatively mapped the lifetimes of various signal transducer and activator of transcription complexes by monitoring dissociation from the surface and defined their stoichiometry on the single molecule level. |
| Databáze: | OpenAIRE |
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