Flor P J, Lindauer K, Püttner I, Rüegg D, Lukic S, Knöpfel T, Kuhn R
Department of Molecular and Cellular Biology, CNS Research, Ciba, Basle, Switzerland.
Eur J Neurosci. 1995 Apr 1;7(4):622-9. doi: 10.1111/j.1460-9568.1995.tb00666.x.
A cDNA encoding the human metabotropic glutamate receptor type 2 (hmGluR2) was isolated from human brain cDNA libraries by cross-hybridization with rat mGluR2 probes. The deduced amino acid sequence of the human mGluR2 receptor consists of 872 residues and shows a sequence identity of 97% to the amino acid sequence of rat mGluR2. Northern blot analyses showed that hmGluR2 is widely expressed in different regions of the adult brain as well as in fetal human brain. Genomic Southern blotting localized the mGluR2 gene to human chromosome 3. Chinese hamster ovary (CHO) cells stably transfected with the cloned hmGluR2 cDNA exhibit agonist induced depression of forskolin-stimulated cAMP accumulation. A direct comparison of CHO cells stably expressing human and rat mGluR2 with five agonists revealed the same rank order of potency [(2S,3S,4S)-alpha-(carboxycyclopropyl)-glycine >> (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid = L-glutamate >> quisqualate = L-2-amino-4-phosphonobutyric acid] and similar EC50 values for both homologous receptors. (R,S)-alpha-methyl-4-carboxyphenylglycine, a reported antagonist at some mGluR subtypes, reduced the depression of forskolin-induced cAMP accumulation by (1S,3R)-ACPD in both human and rat mGluR2.
通过与大鼠mGluR2探针进行交叉杂交，从人脑海绵体cDNA文库中分离出编码人代谢型谷氨酸受体2（hmGluR2）的cDNA。推导的人mGluR2受体氨基酸序列由872个残基组成，与大鼠mGluR2的氨基酸序列显示出97%的序列同一性。Northern印迹分析表明，hmGluR2在成人大脑的不同区域以及胎儿人脑中广泛表达。基因组Southern印迹将mGluR2基因定位到人类染色体3上。用克隆的hmGluR2 cDNA稳定转染的中国仓鼠卵巢（CHO）细胞表现出激动剂诱导的福司可林刺激的cAMP积累的抑制。对稳定表达人和大鼠mGluR2的CHO细胞与五种激动剂进行直接比较，发现两种同源受体的效力顺序相同[（2S，3S，4S）-α-（羧基环丙基）-甘氨酸 >> （1S，3R）-1-氨基环戊烷-1，3-二羧酸 = L-谷氨酸 >> 喹啉酸 = L-2-氨基-4-膦酰丁酸]，并且EC50值相似。（R，S）-α-甲基-4-羧基苯基甘氨酸是一种报道的某些mGluR亚型的拮抗剂，它在人和大鼠mGluR2中均降低了（1S，3R）-ACPD对福司可林诱导的cAMP积累的抑制作用。
