Functional expression of the plant K+ channel KAT1 in insect cells

FEBS Letters
I MartenR Hedrich

Abstract

Following the biophysical analysis of plant K+ channels in their natural environment, three members from the green branch of the evolutionary tree of life KAT1, AKT1, and KST1 have recently been identified on the molecular level. Among them, we focussed on the expression and characterization of the Arabidopsis thaliana K+ channel KAT1 in the insect cell line Sf9. The infection of Sf9 cells with KAT1-recombinant baculovirus resulted in functional expression of KAT1 channels, which was monitored by inward-rectifying, K+-selective (impermeable to Na+ and even NH4+) ionic conductance in whole-cell patch-clamp recordings. A voltage threshold as low as -60 to -80mV for voltage activation compared to other plant inward rectifiers in vivo, and to in vitro expression of KAT1 in Xenopus oocytes or yeast, may be indicative for channel modulation by the expression system. A rise in cytoplasmic Ca2+ concentration (up to 1 mM), a regulator of the inward rectifier in Vicia faba guard cells, did not modify the voltage dependence of KAT1 in Sf9 cells. The access to channel function on one side and channel protein on the other make Sf9 cells a suitable heterologous system for studies on the biophysical properties, post-traditional modification a...Continue Reading

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Citations

Jul 25, 2000·Biochemical and Biophysical Research Communications·I SzabòG M Giacometti
Jun 24, 2006·The Journal of Membrane Biology·F Gambale, N Uozumi
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Jan 8, 2014·Journal of Integrative Plant Biology·Eri Adams, Ryoung Shin
May 6, 2019·International Journal of Molecular Sciences·Antonella LocascioLynne Yenush
Dec 10, 2019·Plant Communications·Ren-Jie TangSheng Luan

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