Rouyer-Fessard C, Augeron C, Grasset E, Maoret J J, Laboisse C L, Laburthe M
Unité de Recherche sur la Différenciation et la Neuroendocrinologie des Cellules Digestives, INSERM U178, Villejuif, France.
Experientia. 1989 Dec 1;45(11-12):1102-5. doi: 10.1007/BF01950169.
At the maximally effective concentration of 10 nM, VIP induced a marked (12.5-fold stimulation above basal), and sustained increase in short circuit current in the human intestinal epithelial cell line Cl.19A grown on permeable filters and placed in Ussing chambers. Half-maximal increase of Isc was observed for 0.1 nM VIP. This was well correlated with the VIP-stimulated adenylate cyclase activity (ED50:0.07 nM). Binding studies using 125I-VIP indicated that Cl.19A cells express a peptide-specific VIP receptor with a dissociation constant of 0.07 nM. Covalent labeling of receptors followed by SDS-PAGE analysis of membrane proteins resulted in the identification of a 63,000 dalton binding protein in Cl.19A cells.
在10 nM的最大有效浓度下,血管活性肠肽(VIP)使生长在可渗透滤膜上并置于尤斯灌流小室中的人肠上皮细胞系Cl.19A的短路电流显著增加(比基础值高12.5倍),且持续升高。对于0.1 nM的VIP,观察到短路电流(Isc)增加至最大值的一半。这与VIP刺激的腺苷酸环化酶活性(半数有效剂量:0.07 nM)密切相关。使用125I-VIP的结合研究表明,Cl.19A细胞表达一种肽特异性VIP受体,其解离常数为0.07 nM。对受体进行共价标记,然后对膜蛋白进行SDS-PAGE分析,结果在Cl.19A细胞中鉴定出一种63,000道尔顿的结合蛋白。