Immunofluorescence-based assay to identify modulators of the number of plasma membrane KCa3.1 channels

被引：7

作者：

Balut, Corina M. ^{[1
]}

Gao, Yajuan ^{[1
]}

Luke, Cliff ^{[2
]}

Devor, Daniel C. ^{[1
]}

机构：

[1] Univ Pittsburgh, Dept Cell Biol & Physiol, Pittsburgh, PA 15261 USA

[2] Univ Pittsburgh, Dept Pediat, Childrens Hosp Pittsburgh, Pittsburgh, PA 15224 USA

来源：

FUTURE MEDICINAL CHEMISTRY | 2010年 / 2卷 / 05期

关键词：

CA2+-ACTIVATED K+ CHANNELS; ACTIVATED POTASSIUM CHANNELS; SMALL-CONDUCTANCE; CHLORIDE SECRETION; CL-SECRETION; INTERMEDIATE; ENDOTHELIUM; DISEASE; MICE; DIFFERENTIATION;

D O I：

10.4155/FMC.10.182

中图分类号：

R914 [药物化学];

学科分类号：

100701 ;

摘要：

Background: Intermediate conductance Ca(2+)-dependent K(+) channels (KCa3.1) have been proposed as therapeutic targets for numerous diseases. We recently characterized the endocytic fate of these channels; leading to the possibility that this can be pharmacologically manipulated, thereby altering the number of channels (N) at the plasma membrane. Results & discussion: We demonstrate that plasma membrane-localized KCa3.1 can be rapidly (10 min) tagged with a fluorophore using a combination of a biotin ligase (BirA) acceptor peptide-tagged channel and an ER-localized BirA. Endocytosis of KCa3.1 was quantified using a 96-well plate format, demonstrating that the ubiquitin-activating enzyme El inhibitor UBEI-41, blocks the endocytosis of KCa3.1. Conclusion: We describe a novel method for identifying modulators of KCa endocytosis and demonstrate this can be used to modulate N at the plasma membrane. It is anticipated that altering N will provide novel therapeutic strategies for targeting these channels in disease.

引用

页码：707 / 713

页数：7

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