Ligand-directed labeling of opioid receptors for covalent attachment of fluorophores or small-molecule probes

Hayden Adoff; Victoria S. Halls; Emily Holland; Braden Lobingier; Seksiri Arttamangkul

doi:10.1016/j.xpro.2023.102231

Ligand-directed labeling of opioid receptors for covalent attachment of fluorophores or small-molecule probes

Hayden Adoff, Victoria S. Halls, Emily Holland, Braden Lobingier, Seksiri Arttamangkul

Research output: Contribution to journal › Article › peer-review

Abstract

This protocol describes endogenous labeling of opioid receptors (ORs) using a ligand-directed reagent, naltrexamine-acylimidazole compounds (NAI-X). NAI acts by guiding and permanently tagging a small-molecule reporter (X)—such as fluorophores or biotin—to ORs. Here we detail syntheses and uses of NAI-X for OR visualization and functional studies. The NAI-X compounds overcome long-standing challenges in mapping and tracking endogenous ORs as the labeling can be done in situ with live tissues or cultured cells. For complete details on the use and execution of this protocol, please refer to Arttamangkul et al.¹^,²

Original language	English (US)
Article number	102231
Journal	STAR Protocols
Volume	4
Issue number	2
DOIs	https://doi.org/10.1016/j.xpro.2023.102231
State	Published - Jun 16 2023

Keywords

Cell Biology
Cell culture
Chemistry
Flow Cytometry/Mass Cytometry
Microscopy
Molecular/Chemical Probes
Neuroscience

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

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10.1016/j.xpro.2023.102231

Cite this

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title = "Ligand-directed labeling of opioid receptors for covalent attachment of fluorophores or small-molecule probes",

abstract = "This protocol describes endogenous labeling of opioid receptors (ORs) using a ligand-directed reagent, naltrexamine-acylimidazole compounds (NAI-X). NAI acts by guiding and permanently tagging a small-molecule reporter (X)—such as fluorophores or biotin—to ORs. Here we detail syntheses and uses of NAI-X for OR visualization and functional studies. The NAI-X compounds overcome long-standing challenges in mapping and tracking endogenous ORs as the labeling can be done in situ with live tissues or cultured cells. For complete details on the use and execution of this protocol, please refer to Arttamangkul et al.1,2",

keywords = "Cell Biology, Cell culture, Chemistry, Flow Cytometry/Mass Cytometry, Microscopy, Molecular/Chemical Probes, Neuroscience",

author = "Hayden Adoff and Halls, {Victoria S.} and Emily Holland and Braden Lobingier and Seksiri Arttamangkul",

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doi = "10.1016/j.xpro.2023.102231",

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AU - Adoff, Hayden

AU - Halls, Victoria S.

AU - Holland, Emily

AU - Lobingier, Braden

AU - Arttamangkul, Seksiri

PY - 2023/6/16

Y1 - 2023/6/16

N2 - This protocol describes endogenous labeling of opioid receptors (ORs) using a ligand-directed reagent, naltrexamine-acylimidazole compounds (NAI-X). NAI acts by guiding and permanently tagging a small-molecule reporter (X)—such as fluorophores or biotin—to ORs. Here we detail syntheses and uses of NAI-X for OR visualization and functional studies. The NAI-X compounds overcome long-standing challenges in mapping and tracking endogenous ORs as the labeling can be done in situ with live tissues or cultured cells. For complete details on the use and execution of this protocol, please refer to Arttamangkul et al.1,2

AB - This protocol describes endogenous labeling of opioid receptors (ORs) using a ligand-directed reagent, naltrexamine-acylimidazole compounds (NAI-X). NAI acts by guiding and permanently tagging a small-molecule reporter (X)—such as fluorophores or biotin—to ORs. Here we detail syntheses and uses of NAI-X for OR visualization and functional studies. The NAI-X compounds overcome long-standing challenges in mapping and tracking endogenous ORs as the labeling can be done in situ with live tissues or cultured cells. For complete details on the use and execution of this protocol, please refer to Arttamangkul et al.1,2

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KW - Cell culture

KW - Chemistry

KW - Flow Cytometry/Mass Cytometry

KW - Microscopy

KW - Molecular/Chemical Probes

KW - Neuroscience

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Ligand-directed labeling of opioid receptors for covalent attachment of fluorophores or small-molecule probes

Abstract

Keywords

ASJC Scopus subject areas

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