title: Single Cell GFP-Trap Reveals Stoichiometry and Dynamics of Cytosolic Protein Complexes.
abstract: 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.
authors.affiliation: Department of Biology, University of Osnabrück, 49076 Osnabrück, Germany.
authors.forename: Tim
authors.initials: T
authors.lastname: Wedeking
authors.affiliation: Department of Biology, University of Osnabrück, 49076 Osnabrück, Germany.
authors.forename: Sara
authors.initials: S
authors.lastname: Löchte
authors.affiliation: Department of Biology, University of Osnabrück, 49076 Osnabrück, Germany.
authors.forename: Christian P
authors.initials: CP
authors.lastname: Richter
authors.affiliation: Department of Biology, University of Osnabrück, 49076 Osnabrück, Germany.
authors.forename: Maniraj
authors.initials: M
authors.lastname: Bhagawati
authors.affiliation: Department of Biology, University of Osnabrück, 49076 Osnabrück, Germany.
authors.forename: Jacob
authors.initials: J
authors.lastname: Piehler
authors.affiliation: Department of Biology, University of Osnabrück, 49076 Osnabrück, Germany.
authors.forename: Changjiang
authors.initials: C
authors.lastname: You
mesh_terms: D003600:Cytosol / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D059010:Single-Cell Analysis; D013499:Surface Properties
chemical_list: D046912:Multiprotein Complexes
keywords: Protein−protein interaction; functional surface micropatterning; nanobody; signal transducer and activator of transcription; single molecule imaging