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Molecular Identification of First Putative Aquaporins in Snails

机译:蜗牛中首推的水通道蛋白的分子鉴定

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摘要

Aquaporins (AQPs), also known as water channel proteins, are members of a large protein family termed Major Intrinsic Proteins (MIP). The mammalian AQPs have been most comprehensively described, while knowledge about AQPs in invertebrates is limited mainly to insects. Not a single AQP protein has been described in snails to date. Consequently, we decided to search for the proteins in gastropod representatives, namely Lymnaea stagnalis, Catascopia occulta, and Stagnicola palustris (Mollusca; Gastropoda; Pulmonata; Lymnaeidae). Using the molecular approach, we identified L. stagnalis, C. occulta,and S. palustris open reading frames (ORFs) showing homology to AQP genes available in GenBank database, and characterized the encoded proteins, referred to as LsAQP1, CoAQP1, and SpAQP1, respectively. The putative snail aquaporins contain 299 amino acids, have a molecular mass of about 32 kDa, display the general AQP topology and three-dimensional structure congruent with orthodox AQPs, i.e., water-specific ones. Due to high levels of similarity in their characteristics, LsAQP1 was chosen for further studies, as the obtained results were supposed to be applicable for CoAQP1 and SpAQP1. Expression analysis revealed the presence of LsAQP1 transcript in the digestive tract, the cerebral ganglia, the kidney, the reproductive system, and the foot, suggesting that LsAQP1 as well as CoAQP1 and SpAQP1 are ubiquitous proteins and may play important roles in many essential water transport processes. The role appears to be confirmed by results of the yeast growth complementation assay pointing at functionality of LsAQP1. Thus, the obtained results support the AQP expression in gastropod tissues for the first time.Electronic supplementary materialThe online version of this article (doi:10.1007/s00232-014-9629-0) contains supplementary material, which is available to authorized users.
机译:水通道蛋白(AQP),也称为水通道蛋白,是被称为主要内在蛋白(MIP)的大型蛋白家族的成员。已经最全面地描述了哺乳动物的AQP,而有关无脊椎动物中AQP的知识主要限于昆虫。迄今为止,蜗牛中还没有描述一种单一的AQP蛋白。因此,我们决定在腹足纲动物的代表中搜索蛋白质,即腹股沟狼蛛,隐匿性卡塔索皮亚氏菌和鼠尾草(软体动物;腹足纲;腹足纲;肺菌; Ly科)。使用分子方法,我们鉴定出了葡萄球菌,隐球菌和pal.ris的开放阅读框(ORF),它们与GenBank数据库中的AQP基因具有同源性,并鉴定了编码的蛋白,称为LsAQP1,CoAQP1和SpAQP1 , 分别。假定的蜗牛水通道蛋白含有299个氨基酸,分子量约为32 kDa,显示出一般的AQP拓扑结构和与正统AQP(即水特异性AQP)相一致的三维结构。由于它们的特性高度相似,因此选择LsAQP1进行进一步研究,因为获得的结果被认为适用于CoAQP1和SpAQP1。表达分析表明,LsAQP1转录本存在于消化道,脑神经节,肾脏,生殖系统和足部,提示LsAQP1以及CoAQP1和SpAQP1是普遍存在的蛋白质,可能在许多必需的水运输中起重要作用流程。指向LsAQP1的功能的酵母生长互补分析结果似乎证实了这一作用。因此,获得的结果首次支持腹足动物组织中的AQP表达。电子补充材料本文的在线版本(doi:10.1007 / s00232-014-9629-0)包含补充材料,授权用户可以使用。

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