Visualization of agonist-induced sequestration and down-regulation of a green fluorescent protein-tagged β2-adrenergic receptor

被引：151

作者：

Kallal, L ^{[1
]}

Gagnon, AW ^{[1
]}

Penn, RB ^{[1
]}

Benovic, JL ^{[1
]}

机构：

[1] Thomas Jefferson Univ, Kimmel Canc Inst, Dept Microbiol & Immunol, Philadelphia, PA 19107 USA

来源：

JOURNAL OF BIOLOGICAL CHEMISTRY | 1998年 / 273卷 / 01期

关键词：

D O I：

10.1074/jbc.273.1.322

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

To date, the visualization of beta(2)-adrenergic receptor (beta(2)AR) trafficking has been largely limited to immunocytochemical analyses of acute internalization events of epitope-tagged receptors in various transfection systems, The development of a beta(2)AR conjugated with green fluorescent protein (beta(2)AR-GFP) provides the opportunity for a more extensive optical analysis of beta(2)AR sequestration, down-regulation, and recycling in cells. Here we demonstrate that stable expression of beta(2)AR-GFP in HeLa cells enables a detailed temporal and spatial analysis of these events, Time dependent colocalization of beta(2)AR-GFP with rhodamine-labeled transferrin and rhodamine-labeled dextran following agonist exposure demonstrates receptor distribution to early endosomes (sequestration) and lysosomes (down-regulation), respectively. The observed temporal distribution of beta(2)AR-GFP was consistent with measures of receptor sequestration and down-regulation generated by radioligand-receptor binding assays. Cells stimulated with different beta-agonists revealed time courses of beta(2)AR-GFP redistribution reflective of the intrinsic activity of each agonist.

引用

页码：322 / 328

页数：7

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