Title

Kinetics of G-Protein Coupled Receptor Endosomal Trafficking Pathways Revealed By Single Quantum Dots

Abstract

G-protein-coupled receptors (GPCRs) are the largest protein superfamily in the human genome; they comprise 30% of current drug targets and regulate diverse cellular signaling responses. The role of endosomal trafficking in GPCR signaling regulation is gaining substantial consideration. However, this process remains difficult to study due to the inability to distinguish among many individual receptors, simultaneously trafficking within multiple endosomal pathways. Here we show accurate measurement of the internalization and endosomal trafficking of single groups of serotonin (5-hydroxytryptamine, 5-HT) receptors using single quantum dot (QD) probes and quantitative colocalization. We demonstrate that the presence of a QD tag does not interfere with 5-HT receptor internalization or endosomal recycling. Direct measurements show simultaneous trafficking of the 5-HT1A receptor in two distinct endosomal recycling pathways. Single-molecule imaging of endosomal trafficking will significantly impact the understanding of cellular signaling and provide powerful tools to elucidate the actions of GPCR-targeted therapeutics.

Department(s)

Chemistry

Document Type

Article

DOI

https://doi.org/10.1073/pnas.1013763107

Rights Information

This article is freely available online through the PNAS Open Access option.

Keywords

serotonin, single-molecule imaging, endocytosis, nanoparticle, endosome

Publication Date

2010

Journal Title

Proceedings of the National Academy of Sciences

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