Cellular and subcellular distribution of polycystin-2, the protein product of the PKD2 gene
Cellular and subcellular distribution of polycystin-2, the protein product of the PKD2 gene
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DOI：
10.1681/asn.v115814
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发表时间：
2000-05-01
影响因子：
13.6
通讯作者：
Sandford, R
中科院分区：
文献类型：
--
作者：
Foggensteiner, L;Bevan, AP;Sandford, R
PKD 1和PKD 2基因突变分别占常染色体显性多囊肾病病例的85%和15%。多囊蛋白-2，PKD 2基因的产物，被预测为与电压激活的Ca 2+通道家族具有同源性的整合膜蛋白。体外研究表明，它可能与多囊蛋白-1，PKD 1基因产物，通过卷曲螺旋结构域存在于其C-末端结构域相互作用。在这项研究中，多囊蛋白-2的细胞和亚细胞分布的定义和比较多囊蛋白-1。一组兔多克隆抗血清针对多囊蛋白-2，并显示识别一个单一的带一致的多囊蛋白-2在多个组织和细胞系的免疫沉淀和蛋白质印迹。人类和小鼠肾组织的免疫染色显示多胞素-2的广泛和发育调节表达，在Henle袢和远曲小管的粗升支中的肾脏中具有最高水平。相比之下，多囊蛋白-1的表达，而本地化到相同的管段，是最高的集合管。免疫组织化学染色和免疫荧光显微镜定位多囊蛋白-2的肾小管上皮细胞的基底侧质膜与多囊蛋白-1的交界定位相比。多囊蛋白-1和多囊蛋白-2在发育、细胞和亚细胞表达方面的差异表明，除了通过其C末端进行潜在的体内相互作用外，它们还可能能够彼此独立地发挥作用。
Mutations in the PKD1 and PKD2 genes account for 85 and 15% of cases of autosomal dominant polycystic kidney disease, respectively. Polycystin-2, the product of the PKD2 gene, is predicted to be an integral membrane protein with homology to a family of voltage-activated Ca2+ channels. In vitro studies suggest that it may interact with polycystin-1, the PKD1 gene product, via coiled-coil domains present in their C-terminal domains. In this study, the cellular and subcellular distribution of polycystin-2 is defined and compared with polycystin-1. A panel of rabbit polyclonal antisera was raised against polycystin-2 and shown to recognize a single band consistent with polycystin-2 in multiple tissues and cell lines by immunoprecipitation and Western blotting. Immunostaining of human and murine renal tissues demonstrated widespread and developmentally regulated expression of polycytin-2, with highest levels in the kidney in the thick ascending limbs of the loop of Henle and the distal convoluted tubule. In contrast, polycystin-1 expression, while localizing to the same tubular segments, was highest in the collecting ducts. Immunohistochemical staining and immunofluorescence microscopy localized polycystin-2 to the basolateral plasma membrane of kidney tubular epithelial cells compared with the junctional localization of polycystin-1. Differences in the developmental, cellular, and subcellular expression of polycystin-1 and polycystin-2 suggest that they may be able to function independently of each other in addition to a potential in vivo interaction via their C-termini.