Covalent quantum dot receptor linkage via the acyl carrier protein for single-molecule tracking, internalization, and trafficking studies.
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.
Zelman-Femiak
Monika
M
Bio-Imaging Center, Rudolf Virchow Center, University of Würzburg, Würzburg, Germany.
Wang
Kun
K
Gromova
Kira V
KV
Knaus
Petra
P
Harms
Gregory S
GS
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*
D000213:Acyl Carrier Protein; D044168:Receptor, Parathyroid Hormone, Type 1; D011956:Receptors, Cell Surface; D052006:Bone Morphogenetic Protein Receptors, Type II