Howard A, Legon S, Walters J R
Department of Medicine, Royal Postgraduate Medical School, London, United Kingdom.
Am J Physiol. 1993 Nov;265(5 Pt 1):G917-25. doi: 10.1152/ajpgi.1993.265.5.G917.
The intestinal basolateral membrane Ca(2+)-transporting adenosinetriphosphatase is the energy-dependent step in the absorption of dietary Ca2+ by the vitamin D-dependent transcellular pathway. Multiple plasma membrane Ca(2+)-pump isoforms are produced from four genes (PMCA1 to 4) and alternative mRNA splicing. We have studied which isoforms are detectable in adult human and rat gastrointestinal tissues by polymerase chain reaction (PCR) amplification, sequencing, and blotting. PMCA1 was the predominant gene product amplified from human small intestinal mucosa, although a minor additional variant lacking the exon at splice site B was detected, which resembled that described for PMCA4. Of the variants described at site C, only the shortest transcript of PMCA1 was amplified; both previously described forms of PMCA4 were found, particularly in colon where PMCA4 predominated. From rat intestinal cDNA, mixed primer PCR amplified PMCA1 and a novel sequence, the rat PMCA4 homologue, which was expressed in many tissues including small intestinal muscle and colon. However, PMCA1 was overwhelmingly predominant in the mucosa of the small intestine, being most abundant in duodenum. These results suggest the involvement of the Ca(2+)-pump isoform PMCA1b in intestinal Ca2+ absorption.
肠道基底外侧膜钙离子转运三磷酸腺苷酶是维生素D依赖的跨细胞途径吸收膳食钙过程中的能量依赖步骤。多种质膜钙离子泵亚型由四个基因(PMCA1至4)通过可变mRNA剪接产生。我们通过聚合酶链反应(PCR)扩增、测序和印迹法研究了在成年人类和大鼠胃肠道组织中可检测到哪些亚型。PMCA1是从小肠黏膜中扩增出的主要基因产物,不过还检测到一种在剪接位点B缺少外显子的次要变体,它类似于PMCA4中描述的变体。在C位点描述的变体中,仅扩增出了PMCA1最短的转录本;发现了两种先前描述的PMCA4形式,尤其是在PMCA4占主导的结肠中。从大鼠肠道cDNA中,混合引物PCR扩增出了PMCA1和一个新序列,即大鼠PMCA4同源物,它在包括小肠肌层和结肠在内的许多组织中表达。然而,PMCA1在小肠黏膜中占绝对优势,在十二指肠中最为丰富。这些结果表明钙离子泵亚型PMCA1b参与了肠道钙吸收。