title: Artificial Formation and Tuning of Glycoprotein Networks on Live Cell Membranes: A Single-Molecule Tracking Study.
abstract: We present a method to artificially induce network formation of membrane glycoproteins and show the precise tuning of their interconnection on living cells. For this, membrane glycans are first metabolically labeled with azido sugars and then tagged with biotin by copper-free click chemistry. Finally, these biotin-tagged membrane proteins are interconnected with streptavidin (SA) to form an artificial protein network in analogy to a lectin-induced lattice. The degree of network formation can be controlled by the concentration of SA, its valency, and the concentration of biotin on membrane proteins. This was verified by investigation of the spatiotemporal dynamics of the SA-protein networks employing single-molecule tracking. It was also proven that this network formation strongly influences the biologically relevant process of endocytosis as it is known from natural lattices on the cell surface.
authors.affiliation: Department of Physical Chemistry, Ludwig Maximilian University of Munich, Butenandtstr. 11, 81377, Munich, Germany.
authors.forename: Leonhard
authors.initials: L
authors.lastname: Möckl
authors.affiliation: Otto Diels Institute of Organic Chemistry, Christiana Albertina University of Kiel, Otto-Hahn-Platz 3-4, 24098, Kiel, Germany.
authors.forename: Thisbe K
authors.initials: TK
authors.lastname: Lindhorst
authors.affiliation: Department of Physical Chemistry, Ludwig Maximilian University of Munich, Butenandtstr. 11, 81377, Munich, Germany. Christoph.Braeuchle@cup.lmu.de.
authors.forename: Christoph
authors.initials: C
authors.lastname: Bräuchle
mesh_terms: D001386:Azides / Q000737:chemistry; D001710:Biotin / Q000737:chemistry; D002232:Carbocyanines / Q000737:chemistry; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D057930:Click Chemistry; D004058:Diffusion; D005456:Fluorescent Dyes / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D019809:Streptavidin / Q000737:chemistry
chemical_list: C569686:Alexa Fluor 647; D001386:Azides; D002232:Carbocyanines; D005456:Fluorescent Dyes; D006023:Glycoproteins; D008565:Membrane Proteins; D001710:Biotin; D019809:Streptavidin
keywords: artificial protein networks; endocytosis; glycoprotein engineering; membrane proteins; single-molecule tracking