Single Cell GFP-Trap Reveals Stoichiometry and Dynamics of Cytosolic Protein Complexes
| Autor(en): | Wedeking, Tim Loechte, Sara Richter, Christian P. Bhagawati, Maniraj Piehier, Jacob You, Changjiang |
Stichwörter: | ASSOCIATION; Chemistry; Chemistry, Multidisciplinary; Chemistry, Physical; functional surface micropatterning; LIVE CELLS; LIVING CELLS; MASS-SPECTROMETRY; Materials Science; Materials Science, Multidisciplinary; MOLECULE PULL-DOWN; MONOFUNCTIONALIZATION; nanobody; Nanoscience & Nanotechnology; Physics; Physics, Applied; Physics, Condensed Matter; PLASMA-MEMBRANE; PREASSOCIATION; Protein-protein interaction; Science & Technology - Other Topics; signal transducer and activator of transcription; SIGNALING COMPLEXES; single molecule imaging; STAT1 | Erscheinungsdatum: | 2015 | Herausgeber: | AMER CHEMICAL SOC | Journal: | NANO LETTERS | Volumen: | 15 | Ausgabe: | 5 | Startseite: | 3610 | Seitenende: | 3615 | Zusammenfassung: | 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. |
ISSN: | 15306984 | DOI: | 10.1021/acs.nanolett.5b01153 |
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geprüft am 14.05.2024
