title: Covalent quantum dot receptor linkage via the acyl carrier protein for single-molecule tracking, internalization, and trafficking studies.
abstract: Here we describe a labeling technique for the covalent linkage of quantum dots to transmembrane receptors for single-molecule tracking. Our method combines the acyl carrier protein (ACP) technique with coenzyme A (CoA)-functionalized quantum dots to covalently attach quantum dots to ACP fusions of receptor proteins. The advantages of this approach include: (i) the use of a smaller attachment linker than in many other quantum dot-labeling systems; (ii) the ability to achieve a reliable 1:1 fluorophore-to-receptor labeling stoichiometry; (iii) the specificity of the method; and (iv) the covalent nature of the quantum dot linkage. We demonstrate the general suitability of this technique in single-molecule tracking, internalization, and trafficking studies by imaging two different transmembrane receptors in living cells.
authors.affiliation: Bio-Imaging Center, Rudolf Virchow Center, University of Würzburg, Würzburg, Germany.
authors.forename: Monika
authors.initials: M
authors.lastname: Zelman-Femiak
authors.forename: Kun
authors.initials: K
authors.lastname: Wang
authors.forename: Kira V
authors.initials: KV
authors.lastname: Gromova
authors.forename: Petra
authors.initials: P
authors.lastname: Knaus
authors.forename: Gregory S
authors.initials: GS
authors.lastname: Harms
mesh_terms: D000213:Acyl Carrier Protein / Q000378:metabolism*; D052006:Bone Morphogenetic Protein Receptors, Type II / Q000378:metabolism; D002470:Cell Survival; D004705:Endocytosis; D057809:HEK293 Cells; D006801:Humans; D021381:Protein Transport; D045663:Quantum Dots; D044168:Receptor, Parathyroid Hormone, Type 1 / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D000213:Acyl Carrier Protein; D044168:Receptor, Parathyroid Hormone, Type 1; D011956:Receptors, Cell Surface; D052006:Bone Morphogenetic Protein Receptors, Type II