Ion permeation and selectivity of wild-type recombinant rat CNG (rOCNC1) channels expressed in HEK293 cells

被引:5
|
作者
Qu, W
Zhu, XO
Moorhouse, AJ
Bieri, S
Cunningham, AM
Barry, PH [1 ]
机构
[1] Univ New S Wales, Sch Physiol & Pharmacol, Sydney, NSW 2052, Australia
[2] Garvan Inst Med Res, Div Neurobiol, Sydney, NSW 2010, Australia
[3] Univ New S Wales, Sch Paediat, Sydney, NSW 2052, Australia
来源
JOURNAL OF MEMBRANE BIOLOGY | 2000年 / 178卷 / 02期
关键词
recombinant rat cyclic nucleotide-gated; channel (rOCNC1); selectivity; permeation; cation; patch clamp;
D O I
10.1007/s002320010021
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
The permeation properties of adenosine 3', 5'-cyclic monophosphate (cAMP)-activated recombinant rat olfactory cyclic nucleotide-gated channels (rOCNC1) in human embryonic kidney (HEK 293) cells were investigated using inside-out excised membrane parches. The relative permeability of these rOCNC1 channels to monovalent alkali cations and organic cations was determined from measurements of the changes in reversal potential upon replacing sodium in the bathing solution with different test cations. The permeability ratio of Cl- relative to Na+ (P-Cl/P-Na) was about 0.14, confirming that these channels are mainly permeable to cations. The se sequence of relative permeabilities of monovalent alkali metal ions in these channels was P-Na greater than or equal to P-K > P-Li > P-Cs greater than or equal to P-Rb, which closely corresponds to a high-strength field sequence as previously determined for native rat olfactory receptor neurons (ORNs). The permeability sequence for organic cations relative to sodium was P-NH3OH > P-NH4 > P-Na > P-Tris > P-Choline > P-TEA again in good agreement with previous permeability ratios obtained in native rat ORNs. Single-channel conductance sequences agreed surprisingly well with permeability sequences. These conductance measurements also indicated that, even in asymmetric bi-ionic cation solutions, the conductance was somewhat independent of current direction and dependent on the composition of both solutions. These results indicate that the permeability properties of rOCNC1 channels are similar to those of native rat CNG channels, and provide a suitable reference point for exploring the molecular basis of ion selectivity in recombinant rOCNC1 channels using site-directed mutagenesis.
引用
收藏
页码:137 / 150
页数:14
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