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首页> 外文期刊>Neurochemistry International: The International Journal for the Rapid Publication of Critical Reviews, Preliminary and Original Research Communications in Neurochemistry >Regulation of the Na(+)-coupled glutamate transporter EAAT3 by PIKfyve.
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Regulation of the Na(+)-coupled glutamate transporter EAAT3 by PIKfyve.

机译：PIKfyve对Na（+）偶联的谷氨酸转运蛋白EAAT3的调节。

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摘要

The Na(+), glutamate cotransporter EAAT3 is expressed in a wide variety of tissues. It accomplishes transepithelial transport and the cellular uptake of acidic amino acids. Regulation of EAAT3 activity involves a signaling cascade including the phosphatidylinositol-3 (PI3)-kinase, the phosphoinositide dependent kinase PDK1, and the serum and glucocorticoid inducible kinase SGK1. Targets of SGK1include the mammalian phosphatidylinositol-3-phosphate-5-kinase PIKfyve (PIP5K3). The present experiments explored whether PIKfyve participates in the regulation of EAAT3 activity. To this end,EAAT3 was expressed in Xenopus oocytes with or without SGK1 and/or PIKfyve and glutamate-induced current (I(glu)) determined by dual electrode voltage clamp. In Xenopus oocytes expressing EAAT3 but not in water injected oocytes glutamate induced an inwardly directed I(glu). Coexpression of either, SGK1 orPIKfyve, significantly enhanced I(glu) in EAAT3 expressing oocytes. The increased I(glu) was paralleled by increased EAAT3 protein abundance in the oocyte cell membrane. I(glu) and EAAT3 protein abundance were significantly larger in oocytes coexpressing EAAT3, SGK1 and PIKfyve than in oocytes expressingEAAT3 and either, SGK1 or PIKfyve, alone. Coexpression of the inactive SGK1 mutant (K127N)SGK1 did not significantly alter I(glu) in EAAT3 expressing oocytes and completely reversed the stimulating effect ofPIKfyve coexpression on I(glu). The stimulating effect of PIKfyve on I(glu) was abolished by replacement of the serine by alanine in the SGK consensus sequence ((S318A)PIKfyve). Moreover, additional coexpression of(S318A)PIKfyve significantly blunted I(glu) in Xenopus oocytes coexpressing SGK1 and EAAT3. The observations demonstrate that PIKfyve participates in EAAT3 regulation likely downstream of SGK1.
机译：Na（+），谷氨酸共转运蛋白EAAT3在各种各样的组织中表达。它完成上皮运输和酸性氨基酸的细胞吸收。 EAAT3活性的调节涉及一个信号级联反应，包括磷脂酰肌醇3（PI3）激酶，磷酸肌醇依赖性激酶PDK1以及血清和糖皮质激素诱导的激酶SGK1。 SGK1的目标包括哺乳动物磷脂酰肌醇3-磷酸-5-激酶PIKfyve（PIP5K3）。本实验探讨了PIKfyve是否参与EAAT3活性的调节。为此，EAAT3在有或没有SGK1和/或PIKfyve的爪蟾卵母细胞中表达，并通过双电极电压钳测定谷氨酸诱导的电流（I（glu））。在非洲蟾蜍卵母细胞中表达EAAT3，但在注水卵母细胞中不表达，谷氨酸诱导向内定向的I（glu）。 SGK1或PIKfyve的共表达显着增强了表达EAAT3的卵母细胞中的I（glu）。 I（glu）的增加与卵母细胞细胞膜中EAAT3蛋白丰度的增加平行。共表达EAAT3，SGK1和PIKfyve的卵母细胞中I（glu）和EAAT3蛋白的丰度明显高于单独表达EAAT3和SGK1或PIKfyve的卵母细胞。失活的SGK1突变体（K127N）SGK1的共表达不会显着改变表达EAAT3的卵母细胞中的I（glu），并且完全逆转了PIKfyve共表达对I（glu）的刺激作用。通过在SGK共有序列（（S318A）PIKfyve）中用丙氨酸代替丝氨酸，消除了PIKfyve对I（glu）的刺激作用。此外，（S318A）PIKfyve的其他共表达使共同表达SGK1和EAAT3的爪蟾卵母细胞中的I（glu）明显变钝。观察结果表明PIKfyve可能参与了SGK1下游的EAAT3调控。

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