Huelseweh B, Kohl B, Hentschel H, Kinne R K, Werner A
Max-Planck-Institut für Molekulare Physiologie, Rheinlanddamm 201, 44139 Dortmund, Federal Republic of Germany.
Biochem J. 1998 Jun 1;332 ( Pt 2)(Pt 2):483-9. doi: 10.1042/bj3320483.
The homeostasis of Pi in marine teleosts is maintained by renal Pi secretion as well as by Pi reabsorption. A Na/Pi co-transport system belonging to the NaPi-II protein family is instrumental in tightly controlled renal Pi handling in mammals and fish. We have isolated an NaPi-II related cDNA from winter flounder. It was cloned from a female gonad cDNA library and is 624 bp long. The transcript is expressed in female and male flounder gonads as well as in kidney and intestine, although at very low levels. RNase H digestion experiments revealed an opposite orientation of the transcript with regard to NaPi-II-related mRNA. The anti-sense orientation was confirmed by genomic sequence analysis and Southern blotting. Alluding to the sense transcript, the anti-sense transcript was denoted IPAN. The open reading frame of IPAN encodes a basic protein of 68 amino acid residues. Immunohistochemistry confined the anti-sense related protein, Ipan, to a submembranous compartment of immature oocytes, suggesting a role in oocyte development. In kidney and intestine Ipan is partly co-localized with the Na/Pi co-transporter, implying a regulatory function for the anti-sense protein. However, direct protein-protein interaction could not be established. The existence of a putative open reading frame in other species extends the biological significance of the novel protein.
海洋硬骨鱼体内磷(Pi)的稳态通过肾脏分泌磷以及重吸收磷来维持。属于NaPi-II蛋白家族的钠/磷共转运系统在哺乳动物和鱼类严格调控的肾脏磷处理过程中发挥着重要作用。我们从冬比目鱼中分离出了一个与NaPi-II相关的cDNA。它是从雌性性腺cDNA文库中克隆出来的,长度为624 bp。该转录本在雌性和雄性比目鱼的性腺以及肾脏和肠道中均有表达,不过表达水平很低。核糖核酸酶H消化实验表明,该转录本与NaPi-II相关mRNA的方向相反。通过基因组序列分析和Southern印迹法证实了其反义方向。参照正义转录本,该反义转录本被命名为IPAN。IPAN的开放阅读框编码一个由68个氨基酸残基组成的碱性蛋白。免疫组织化学将与反义相关的蛋白Ipan定位在未成熟卵母细胞的膜下区域,表明其在卵母细胞发育中发挥作用。在肾脏和肠道中,Ipan部分与钠/磷共转运体共定位,这意味着该反义蛋白具有调节功能。然而,无法确定直接的蛋白质-蛋白质相互作用。在其他物种中存在推定的开放阅读框,这扩展了这种新蛋白的生物学意义。