Biotinylated-spiperone ligands for quantum dot labeling of the dopamine D2 receptor in live cell cultures

Ian D Tomlinson; Oleg Kovtun; Tiffany M Crescentini; Sandra J Rosenthal

doi:10.1016/j.bmcl.2019.02.024

Biotinylated-spiperone ligands for quantum dot labeling of the dopamine D2 receptor in live cell cultures

Bioorg Med Chem Lett. 2019 Apr 15;29(8):959-964. doi: 10.1016/j.bmcl.2019.02.024. Epub 2019 Feb 21.

Authors

Ian D Tomlinson¹, Oleg Kovtun¹, Tiffany M Crescentini¹, Sandra J Rosenthal²

Affiliations

¹ Department of Chemistry, Vanderbilt University, Nashville, TN 37232, United States.
² Department of Chemistry, Vanderbilt University, Nashville, TN 37232, United States; Department of Pharmacology, Vanderbilt University, Nashville, TN 37232, United States; Departments of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, TN 37232, United States; Departments of Physics and Astronomy, Vanderbilt University, Nashville, TN 37232, United States; Vanderbilt Institute for Nanoscale Science and Engineering, Vanderbilt University, Nashville, TN 37232, United States; Department of Interdisciplinary Materials Science, Vanderbilt University, Nashville, TN 37232, United States; Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, TN 37232, United States. Electronic address: Sandra.J.Rosenthal@vanderbilt.edu.

PMID: 30808590
DOI: 10.1016/j.bmcl.2019.02.024

Abstract

We have synthesized 3 analogs of the dopamine D2 receptor (D2 DR) antagonist spiperone that can be conjugated to streptavidin-coated quantum dots via a pegylated biotin derivative. Using fluorescent imaging we demonstrate that substitution on the spiro position is tolerated, whilst the length and rigidity of a spacer arm attached to spiperone is important in controlling specific labeling as well as minimizing nonspecific labeling to cells and the surface of cell culture dishes. The ligand with the most rigid linker IDT772 (4) had the best binding profile and had high specific binding to D2 DR expressing HEK-293T cells with low nonspecific binding to plates and HEK-293T cells that lacked the D2 DR.

Keywords: Antagonist; D2 receptor; Fluorescent imaging; Quantum dot; Receptor tracking; Spiperone.

MeSH terms

Biotin / chemistry*
Dopamine D2 Receptor Antagonists / chemistry
HEK293 Cells
Humans
Ligands
Microscopy, Fluorescence
Quantum Dots / chemistry*
Receptors, Dopamine D2 / chemistry
Receptors, Dopamine D2 / genetics
Receptors, Dopamine D2 / metabolism*
Spiperone / chemistry*
Streptavidin / chemistry

Substances

Dopamine D2 Receptor Antagonists
Ligands
Receptors, Dopamine D2
Spiperone
Biotin
Streptavidin