Covalent quantum dot receptor linkage via the acyl carrier protein for single-molecule tracking, internalization, and trafficking studies

Biotechniques. 2010 Aug;49(2):574-9. doi: 10.2144/000113466.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acyl Carrier Protein / metabolism*
  • Bone Morphogenetic Protein Receptors, Type II / metabolism
  • Cell Survival
  • Endocytosis*
  • HEK293 Cells
  • Humans
  • Protein Transport
  • Quantum Dots*
  • Receptor, Parathyroid Hormone, Type 1 / metabolism
  • Receptors, Cell Surface / metabolism*
  • Staining and Labeling / methods*

Substances

  • Acyl Carrier Protein
  • Receptor, Parathyroid Hormone, Type 1
  • Receptors, Cell Surface
  • Bone Morphogenetic Protein Receptors, Type II