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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