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• W2113500592 endingPage "C1525" @default.
• W2113500592 startingPage "C1516" @default.
• W2113500592 abstract "The neuronal K-Cl cotransporter isoform (KCC2) was functionally expressed in human embryonic kidney (HEK-293) cell lines. Two stably transfected HEK-293 cell lines were prepared: one expressing an epitope-tagged KCC2 (KCC2–22T) and another expressing the unaltered KCC2 (KCC2–9). The KCC2–22T cells produced a glycoprotein of ∼150 kDa that was absent from HEK-293 control cells. The 86 Rb influx in both cell lines was significantly greater than untransfected control HEK-293 cells. The KCC2–9 cells displayed a constitutively active 86 Rb influx that could be increased further by 1 mM N-ethylmaleimide (NEM) but not by cell swelling. Both furosemide [inhibition constant ( K i ) ∼25 μM] and bumetanide (K i ∼55 μM) inhibited the NEM-stimulated 86 Rb influx in the KCC2–9 cells. This diuretic-sensitive 86 Rb influx in the KCC2–9 cells, operationally defined as KCC2 mediated, required external Cl − but not external Na + and exhibited a high apparent affinity for external Rb + (K + ) [Michaelis constant ( K m ) = 5.2 ± 0.9 (SE) mM; n = 5] but a low apparent affinity for external Cl − ( K m >50 mM). On the basis of thermodynamic considerations as well as the unique kinetic properties of the KCC2 isoform, it is hypothesized that KCC2 may serve a dual function in neurons: 1) the maintenance of low intracellular Cl − concentration so as to allow Cl − influx via ligand-gated Cl − channels and 2) the buffering of external K + concentration ([K + ] o ) in the brain." @default.
• W2113500592 created "2016-06-24" @default.
• W2113500592 creator A5050619139 @default.
• W2113500592 date "1997-11-01" @default.
• W2113500592 modified "2023-10-14" @default.
• W2113500592 title "Functional characterization of the neuronal-specific K-Cl cotransporter: implications for [K⁺]_oregulation" @default.
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• W2113500592 doi "https://doi.org/10.1152/ajpcell.1997.273.5.c1516" @default.
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