Peptide-labeled quantum dots for imaging GPCRs in whole cells and as single molecules

Min Zhou, Eri Nakatani, Luisa S. Gronenberg, Takahira Tokimoto, Mary J. Wirth, Victor J Hruby, Amber Roberts, Ron Lynch, Indraneel Ghosh

Research output: Contribution to journalArticle

87 Citations (Scopus)

Abstract

We report a robust and practical method for the preparation of water-soluble luminescent quantum dots (QDs) selectively coupled through an amine or thiol linkage to peptide ligands targeted to G-protein coupling receptors (GPCRs) and demonstrate their utility in whole-cell and single-molecule imaging. We utilized a low molecular weight (∼1200 Da) diblock copolymer with acrylic acids as hydrophilic segments and amido-octyl side chains as hydrophobic segments for facile encapsulation of QDs (QD 595 and QD 514) in aqueous solutions. As proof of principle, these QDs were targeted to the human melanocortin receptor (hMCR) by chemoselectively coupling the polymer-coated QDs to either a hexapeptide analog of α-melanocyte stimulating hormone or to the highly potent MT-II ligand containing a unique amine. To label QDs with ligands lacking orthogonal amines, the diblock copolymers were readily modified with water-soluble trioxa-tridecanediamine to incorporate freely available amine functionalities. The amine-functionalized QDs underwent facile reaction with the bifunctional linker NHS-maleimide, allowing for covalent coupling to GPCR-targeted ligands modified with unique cysteines. We demonstrate the utility of these maleimide-functionalized QDs by covalent conjugation to a highly potent Deltorphin-II analog that allowed for selective cell-surface and single-molecule imaging of the human δ-opioid receptor (hDOR).

Original languageEnglish (US)
Pages (from-to)323-332
Number of pages10
JournalBioconjugate Chemistry
Volume18
Issue number2
DOIs
StatePublished - Mar 2007

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Quantum Dots
GTP-Binding Proteins
Peptides
Semiconductor quantum dots
Proteins
Imaging techniques
Molecules
Amines
Ligands
Block copolymers
Melanocortin Receptors
Melanocyte-Stimulating Hormones
Water
Hormones
Opioid Receptors
Encapsulation
Sulfhydryl Compounds
Acrylics
Cysteine
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ASJC Scopus subject areas

  • Chemistry(all)
  • Organic Chemistry
  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Peptide-labeled quantum dots for imaging GPCRs in whole cells and as single molecules. / Zhou, Min; Nakatani, Eri; Gronenberg, Luisa S.; Tokimoto, Takahira; Wirth, Mary J.; Hruby, Victor J; Roberts, Amber; Lynch, Ron; Ghosh, Indraneel.

In: Bioconjugate Chemistry, Vol. 18, No. 2, 03.2007, p. 323-332.

Research output: Contribution to journalArticle

Zhou, Min ; Nakatani, Eri ; Gronenberg, Luisa S. ; Tokimoto, Takahira ; Wirth, Mary J. ; Hruby, Victor J ; Roberts, Amber ; Lynch, Ron ; Ghosh, Indraneel. / Peptide-labeled quantum dots for imaging GPCRs in whole cells and as single molecules. In: Bioconjugate Chemistry. 2007 ; Vol. 18, No. 2. pp. 323-332.
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