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Diffusion and Interaction Dynamics of Individual Membrane Protein Complexes Confined in Micropatterned Polymer‐Supported Membranes
Author(s) -
Waichman Sharon,
Roder Friedrich,
Richter Christian P.,
Birkholz Oliver,
Piehler Jacob
Publication year - 2013
Publication title -
small
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.785
H-Index - 236
eISSN - 1613-6829
pISSN - 1613-6810
DOI - 10.1002/smll.201201530
Subject(s) - fluorescence recovery after photobleaching , membrane , diffusion , transmembrane protein , materials science , polymer , vesicle , photobleaching , biophysics , molecule , vesicle fusion , fluorescence microscope , lipid bilayer fusion , nanotechnology , chemistry , fluorescence , biochemistry , organic chemistry , receptor , physics , quantum mechanics , biology , synaptic vesicle , composite material , thermodynamics
Micropatterned polymer‐supported membranes (PSM) are established as a tool for confining the diffusion of transmembrane proteins for single molecule studies. To this end, a photochemical surface modification with hydrophobic tethers on a PEG polymer brush is implemented for capturing of lipid vesicles and subsequent fusion. Formation of contiguous membranes within micropatterns is confirmed by scanning force microscopy, fluorescence recovery after photobleaching (FRAP), and super‐resolved single‐molecule tracking and localization microscopy. Free diffusion of transmembrane proteins reconstituted into micropatterned PSM is demonstrated by FRAP and by single‐molecule tracking. By exploiting the confinement of diffusion within micropatterned PSM, the diffusion and interaction dynamics of individual transmembrane receptors are quantitatively resolved.