Abstract
BLAP-Tagged Channel Technology: A New Direction Trafficking toward Epithelial Physiology
Highlights
Devor and colleagues (Balut et al, 2010b; Gao et al, 2010), using the biotin-ligase acceptor peptide (BLAP)-technology based on work of Ting, have defined the membrane residence and retrograde trafficking of the Ca2+-activated K+ channels, KCa3.1 and KCa2.3, in human embryonic kidney (HEK293) cells and endothelial cells (HMEC-1)
Gao et al (2010) demonstrated that KCa2.3 had a long membrane residence time (∼13 h) and that the channel was recycled back to the membrane, while KCa3.1 had a short membrane residence time (∼ 90 min), and was destined for the lysosomes (Balut et al, 2010b). Both KCa3.1 and KCa2.3 have been suggested as therapeutic targets in many diseases including endothelial dysfunction and hypertension (Kohler et al, 2010)
Devor and colleagues realized the time of labeling of KCa3.1 must be reduced in order to rapidly assess modulators of channel endocytosis, and for the effectiveness of their procedure to be functionally viable in a 96-well plate format assay approach with future application in a highthroughput scenario
Summary
Devor and colleagues (Balut et al, 2010b; Gao et al, 2010), using the BLAP-technology based on work of Ting, have defined the membrane residence and retrograde trafficking of the Ca2+-activated K+ channels, KCa3.1 and KCa2.3, in human embryonic kidney (HEK293) cells and endothelial cells (HMEC-1). The focus of this Commentary is to highlight recent work of Devor and colleagues (Balut et al, 2010a) in which they developed a novel immunofluorescentbased assay to assess modulators of endocytosis of proteins (e.g., KCa3.1).
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