Involvement of Direct Phosphorylation in the Regulation of the Rat Parotid Na+-K+-2Cl− Cotransporter (*)
Involvement of Direct Phosphorylation in the Regulation of the Rat Parotid Na+-K+-2Cl− Cotransporter (*)
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直接磷酸化参与大鼠腮腺 Na+-K+-2Cl− 协同转运蛋白的调节 (*)
DOI：
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发表时间：
1995
影响因子：
4.8
通讯作者：
R. J. Turner
中科院分区：
文献类型：
--
作者：
A. Tanimura;K. Kurihara;Stephan J. Reshkin;R. J. Turner
我们在大鼠腮腺腺泡中鉴定了一种175 kDa的膜磷蛋白（pp 175），其性质与先前在该组织中表征的Na+-K+-2Cl−协同转运蛋白的功能和生化性质密切相关。pp 175是唯一一种被抗Na +-K+-2Cl−协同转运蛋白抗体免疫沉淀的磷蛋白，也是唯一一种在β-肾上腺素能激动剂异丙肾上腺素短暂（45秒）暴露于腺泡后磷酸化状态发生显著改变的膜蛋白。磷酸肽图谱提供了证据的三个磷酸化位点的pp 175，其中只有一个被标记在响应异丙肾上腺素治疗。异丙肾上腺素对pp 175磷酸化的半数最大效应（约20 nM）与其先前证明的对共转运活性的上调效应非常一致。增加磷酸化的pp 175也被视为以下的腺泡治疗与渗透性cAMP类似物和毛喉素，条件下，也被证明是上调协同转运蛋白。结合我们实验室的早期结果，这些数据提供了强有力的证据，这些药物上调协同转运蛋白是由于蛋白激酶A介导的直接磷酸化。AlF 4 −处理导致共转运活性的上调，与异丙肾上腺素观察到的结果相当（约6倍），导致pp 175磷酸化的类似增加。然而，高渗收缩和治疗与蛋白磷酸酶抑制剂calyculin A，这也上调协同转运蛋白（分别为103倍和106倍）没有引起磷酸化水平的变化。此外，虽然与毒蕈碱激动剂卡巴胆碱的腺泡治疗的结果在一个显着的上调共转运活性和伴随的磷酸化的pp 175，没有磷酸化的pp 175被视为与钙离子动员剂毒胡萝卜素，这是能够完全模仿卡巴胆碱对运输活性的上调作用。总之，这些结果表明，直接磷酸化只是促分泌素诱导的大鼠腮腺Na+-K+-2Cl−协同转运蛋白调节的机制之一。
We identify a 175-kDa membrane phosphoprotein (pp175) in rat parotid acini whose properties correlate well with the Na+-K+-2Cl− cotransporter previously characterized functionally and biochemically in this tissue. pp175 was the only phosphoprotein immunoprecipitated by an anti-Na+-K+-2Cl− cotransporter antibody and the only membrane protein whose phosphorylation state was conspicuously altered after a brief (45-s) exposure of acini to the β-adrenergic agonist isoproterenol. Phosphopeptide mapping provided evidence for three phosphorylation sites on pp175, only one of which was labeled in response to isoproterenol treatment. The half-maximal effect of isoproterenol on phosphorylation of pp175 (≈20 nM) was in excellent agreement with its previously demonstrated up-regulatory effect on cotransport activity. Increased phosphorylation of pp175 was also seen following acinar treatment with a permeant cAMP analogue and with forskolin, conditions that have likewise been shown to up-regulate the cotransporter. Combined with earlier results from our laboratory, these data provide strong evidence that the up-regulation of the cotransporter by these agents is due to direct phosphorylation mediated by protein kinase A. AlF4− treatment, which results in an up-regulation of cotransport activity comparable with that observed with isoproterenol (∼6-fold), caused a similar increase in phosphorylation of pp175. However, hypertonic shrinkage and treatment with the protein phosphatase inhibitor calyculin A, which also up-regulate the cotransporter (∼3-fold and ∼6-fold, respectively) caused no change in the phosphorylation level. Furthermore, although acinar treatment with the muscarinic agonist carbachol results in a dramatic up-regulation of cotransport activity and a concomitant phosphorylation of pp175, no phosphorylation of pp175 was seen with the Ca2+-mobilizing agent thapsigargin, which is able to fully mimic the up-regulatory effect of carbachol on transport activity. Taken together, these results indicate that direct phosphorylation is only one of the mechanisms involved in secretagogue-induced regulation of the rat parotid Na+-K+-2Cl− cotransporter.
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鲨鱼直肠腺的 Na-K-Cl 共转运蛋白。
DOI：
--
发表时间：
1992
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Lytle,C;Forbush3rd,B
通讯作者：
Forbush3rd,B
DOI：
10.1016/0006-291x(89)92189-x
发表时间：
1989-03-31
影响因子：
3.1
作者：
ISHIHARA, H;MARTIN, BL;HARTSHORNE, DJ
通讯作者：
HARTSHORNE, DJ
鲨鱼直肠腺中的 Na-K-Cl 共转运。
DOI：
10.1152/ajpcell.1992.262.4.c1000
发表时间：
1992
期刊：
The American journal of physiology
影响因子：
--
作者：
Forbush3rd,B;Haas,M;Lytle,C
通讯作者：
Lytle,C
禽类盐腺的 Na-K-Cl 协同转运蛋白。
DOI：
--
发表时间：
1992
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Torchia,J;Lytle,C;Pon,DJ;Forbush3rd,B;Sen,AK
通讯作者：
Sen,AK
使用布美他尼类似物对来自鸭红细胞的 150 kDa (Na K Cl) 共转运蛋白进行光亲和标记。
DOI：
10.1016/0005-2736(88)90054-5
发表时间：
1988
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Haas,M;Forbush3rd,B
通讯作者：
Forbush3rd,B