Differential activation of "social" and "solitary" variants of the Caenorhabditis elegans G protein-coupled receptor NPR-1 by its cognate ligand AF9
Differential activation of "social" and "solitary" variants of the Caenorhabditis elegans G protein-coupled receptor NPR-1 by its cognate ligand AF9
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DOI：
10.1074/jbc.m304861200
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发表时间：
2003-09-05
影响因子：
4.8
通讯作者：
Lowery, DE
中科院分区：
文献类型：
--
作者：
Kubiak, TM;Larsen, MJ;Lowery, DE
野生秀丽隐杆线虫分离株的天然变异在NPR-1（一种假定的孤儿神经肽Y样G蛋白偶联受体）中具有Phe-215或瓦尔-215，导致“群居”或“孤独”进食行为（de Bono，M.，和Bargmann，C. I.（1998）Cell 94，679 - 689）。我们确定了一种线虫肽，GLGPR-PLRF-NH 2（AF 9），作为激活哺乳动物细胞中异源表达的克隆NPR-1受体的配体。将细胞培养温度从37 ℃转变为28 ℃，在转染后24小时实施，对于NPR-1的可检测功能表达至关重要。AF 9处理将两种克隆的受体变体与Gi/Go蛋白的激活和cAMP抑制联系起来，从而允许将NPR-1分类为抑制性G蛋白偶联受体。瓦尔-215受体亚型显示出比其Phe-215对应物更高的结合和功能活性。这一发现与体内观察到的由编码瓦尔-215受体形式的npr-1基因对社会进食的更有效抑制相平行，导致分散（孤立）动物。由于神经肽Y与AF 9没有序列同源性，并且在克隆的NPR-1上没有功能活性，我们建议将NPR-1重新命名为AF 9受体，AF 9-R1。
Natural variations of wild Caenorhabditis elegans isolates having either Phe-215 or Val-215 in NPR-1, a putative orphan neuropeptide Y-like G protein-coupled receptor, result in either "social" or "solitary" feeding behaviors ( de Bono, M., and Bargmann, C. I. ( 1998) Cell 94, 679 - 689). We identified a nematode peptide, GLGPR-PLRF-NH2 (AF9), as a ligand activating the cloned NPR-1 receptor heterologously expressed in mammalian cells. Shifting cell culture temperatures from 37 to 28 degreesC, implemented 24 h after transfections, was essential for detectable functional expression of NPR-1. AF9 treatments linked both cloned receptor variants to activation of Gi/Go proteins and cAMP inhibition, thus allowing for classification of NPR-1 as an inhibitory G protein-coupled receptor. The Val-215 receptor isoform displayed higher binding and functional activity than its Phe-215 counterpart. This finding parallels the in vivo observation of a more potent repression of social feeding by the npr-1 gene encoding the Val-215 form of the receptor, resulting in dispersing ( solitary) animals. Since neuropeptide Y shows no sequence homology to AF9 and was functionally inactive at the cloned NPR-1, we propose to rename NPR-1 and refer to it as an AF9 receptor, AF9-R1.