Mulders S M, Bichet D G, Rijss J P, Kamsteeg E J, Arthus M F, Lonergan M, Fujiwara M, Morgan K, Leijendekker R, van der Sluijs P, van Os C H, Deen P M
Department Of Cell Physiology, University of Nijmegen, 6500 HB Nijmegen, The Netherlands.
J Clin Invest. 1998 Jul 1;102(1):57-66. doi: 10.1172/JCI2605.
Mutations in the aquaporin-2 (AQP2) water channel gene cause autosomal recessive nephrogenic diabetes insipidus (NDI). Here we report the first patient with an autosomal dominant form of NDI, which is caused by a G866A transition in the AQP2 gene of one allele, resulting in a E258K substitution in the C-tail of AQP2. To define the molecular cause of NDI in this patient, AQP2-E258K was studied in Xenopus oocytes. In contrast to wild-type AQP2, AQP2-E258K conferred a small increase in water permeability, caused by a reduced expression at the plasma membrane. Coexpression of wild-type AQP2 with AQP2-E258K, but not with an AQP2 mutant in recessive NDI (AQP2-R187C), revealed a dominant-negative effect on the water permeability conferred by wild-type AQP2. The physiologically important phosphorylation of S256 by protein kinase A was not affected by the E258K mutation. Immunoblot and microscopic analyses revealed that AQP2-E258K was, in contrast to AQP2 mutants in recessive NDI, not retarded in the endoplasmic reticulum, but retained in the Golgi compartment. Since AQPs are thought to tetramerize, the retention of AQP2-E258K together with wild-type AQP2 in mixed tetramers in the Golgi compartment is a likely explanation for the dominant inheritance of NDI in this patient.
水通道蛋白2(AQP2)水通道基因的突变会导致常染色体隐性遗传性肾源性尿崩症(NDI)。在此,我们报告了首例常染色体显性形式的NDI患者,其病因是一个等位基因的AQP2基因发生了G866A转换,导致AQP2的C末端出现E258K替代。为了明确该患者NDI的分子病因,我们在非洲爪蟾卵母细胞中对AQP2 - E258K进行了研究。与野生型AQP2不同,AQP2 - E258K使水通透性略有增加,这是由质膜上表达降低所致。野生型AQP2与AQP2 - E258K共表达,但与隐性NDI中的AQP2突变体(AQP2 - R187C)不共表达,结果显示对野生型AQP2赋予的水通透性有显性负效应。蛋白激酶A对S256的重要生理性磷酸化不受E258K突变影响。免疫印迹和显微镜分析显示,与隐性NDI中的AQP2突变体不同,AQP2 - E258K在内质网中未被滞留,而是保留在高尔基体区室。由于水通道蛋白被认为会形成四聚体,AQP2 - E258K与野生型AQP2在高尔基体区室的混合四聚体中共同保留,这可能是该患者NDI显性遗传的原因。