Tunwell R E, Lai F A
Medical Research Council National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, United Kingdom.
J Biol Chem. 1996 Nov 22;271(47):29583-8. doi: 10.1074/jbc.271.47.29583.
An oligonucleotide probe to a conserved 3' region within the three identified ryanodine receptor-calcium release channel isoforms hybridized to a single clone from a rabbit kidney cDNA library. The kidney clone encoded the carboxyl-terminal 338 amino acids within the putative transmembrane domain of the type 2 ryanodine receptor sequence. Reverse transcriptase-polymerase chain reaction with isoform-specific oligonucleotide primers demonstrated the presence of the type 2 ryanodine receptor transcript in rabbit kidney, as well as in a non-excitable cell line, LLC-RK1, derived from rabbit kidney epithelial cells. Amplification by rapid amplification of 5' cDNA ends indicated the kidney type 2 ryanodine receptor transcript extended >7000 base pairs from the stop codon and is therefore not homologous to the short RyR-1 transcript of approximately 2500 base pairs previously observed in rabbit brain. [3H]Ryanodine binding and immunoblot analysis with a type 2 ryanodine receptor-specific antibody demonstrated that the native type 2 ryanodine receptor protein is expressed in the kidney. These observations suggest that the type 2 ryanodine receptor isoform may play a functional role in regulating intracellular calcium homeostasis in non-excitable cells.
一种针对三种已鉴定的兰尼碱受体 - 钙释放通道亚型中保守3'区域的寡核苷酸探针，与兔肾cDNA文库中的一个单一克隆杂交。该肾克隆编码了2型兰尼碱受体序列假定跨膜结构域内的羧基末端338个氨基酸。用亚型特异性寡核苷酸引物进行逆转录 - 聚合酶链反应表明，2型兰尼碱受体转录本存在于兔肾以及源自兔肾上皮细胞的非兴奋性细胞系LLC - RK1中。通过5' cDNA末端快速扩增进行的扩增表明，肾2型兰尼碱受体转录本从终止密码子延伸超过7000个碱基对，因此与先前在兔脑中观察到的约2500个碱基对的短RyR - 1转录本不同源。用2型兰尼碱受体特异性抗体进行的[3H]兰尼碱结合和免疫印迹分析表明，天然2型兰尼碱受体蛋白在肾中表达。这些观察结果表明，2型兰尼碱受体亚型可能在调节非兴奋性细胞内的细胞内钙稳态中发挥功能作用。
