Electrophysiological characterization and ionic stoichiometry of the rat brain K+-dependent Na+/Ca2+ exchanger, NCKX2

被引:53
作者
Dong, H
Light, PE
French, RJ
Lytton, J
机构
[1] Univ Calgary, Hlth Sci Ctr, Dept Biochem & Mol Biol, Calgary, AB T2N 4N1, Canada
[2] Univ Calgary, Dept Physiol & Biophys, Calgary, AB T2N 4N1, Canada
关键词
D O I
10.1074/jbc.M103401200
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
We have recently described a novel K+-dependent Na+/Ca2+ exchanger, NCKX2, that is abundantly expressed in brain neurons (Tsoi, M,, Rhee, IL-H., Bungard, D,, Li, X-F,, Lee, S,-L,, Auer, R, N,, and Lytton, J, (1998) J, Biol, Chem, 273, 4115-4162), The precise role for NCKX2 in neuronal Ca2+ homeostasis is not yet clearly understood but will depend upon the functional properties of the molecule. Here, we have performed whole-cell patch clamp analysis to characterize cation dependences and ion stoichiometry for rat brain NCKX2, heterologously expressed in HEK293 cells. Outward currents generated by reverse NCKX2 exchange depended on external Ca2+ with a K-1/2 of 1.4 or 101 muM without or with 1 mM Mg2+, and on external K+ with a K-1/2 of about 12 or 36 mM with choline or Li+ as counter ion, respectively. Na+ inhibited outward currents with a K-1/2 of about 60 mM, Inward currents generated by forward NCKX2 exchange depended upon external Na+ with a K-1/2 of 30 mM and a Hill coefficient of 2.8. K+ inhibited the inward currents by a maximum of 40%, with a K-1/2 of 2 mM or less, depending upon the conditions. The transport stoichiometry of NCKX2 was determined by observing the change in reversal potential as individual ion gradients were altered. Our data support a stoichiometry for rat brain NCKX2 of 4 Na+:(1 Ca2+ + 1 K+), These findings provide the first electrophysiological characterization of rat brain NCKX2, and the first evidence that a single recombinantly expressed NCKX polypeptide encodes a K+-transporting Na+/Ca2+ exchanger with a transport stoichiometry of 4 Na+:(1 Ca2+ + 1 K+).
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页码:25919 / 25928
页数:10
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