使用新型非肽Y1受体选择性拮抗剂[3H]BIBP3226对SK-N-MC细胞中的神经肽Y受体进行标记。

Labeling of neuropeptide Y receptors in SK-N-MC cells using the novel, nonpeptide Y1 receptor-selective antagonist [3H]BIBP3226.

作者信息

Entzeroth M, Braunger H, Eberlein W, Engel W, Rudolf K, Wienen W, Wieland H A, Willim K D, Doods H N

机构信息

Department of Pharma Research, Dr. Karl Thomae GmbH, Biberach, Germany.

出版信息

Eur J Pharmacol. 1995 May 24;278(3):239-42. doi: 10.1016/0014-2999(95)00161-d.

DOI:10.1016/0014-2999(95)00161-d

PMID:7589160

Abstract

The binding of tritium-labelled BIBP3226, N2-(diphenylacetyl)-N-[(4-hydroxy-phenyl)methyl]-D-arginine amide, to human neuroblastoma SK-N-MC cells was investigated. [3H]BIBP3226 reversibly binds to neuropeptide Y receptors of the Y1 subtype expressed in SK-N-MC cells with a KD of 2.1 +/- 0.3 nM (mean +/- S.E.M., n = 3) and a Bmax of 58,400 +/- 1100 sites/cell. Non-specific binding did not exceed 30% of the total radioactivity bound at KD. In competition experiments [3H]BIBP3226 is concentration-dependently displaced by neuropeptide Y and its peptide analogues with an affinity pattern neuropeptide Y = [Leu31, Pro34]neuropeptide Y >> neuropeptide Y-(18-36). This rank order of potencies is consistent with the interaction of [3H]BIBP3226 with neuropeptide Y receptors of the Y1 subtype. Therefore, [3H]BIBP3226 can be used as selective ligand to study neuropeptide Y Y1 receptors.

摘要

研究了氚标记的BIBP3226（N2-（二苯乙酰基）-N-[(4-羟基苯基)甲基]-D-精氨酸酰胺）与人神经母细胞瘤SK-N-MC细胞的结合情况。[3H]BIBP3226以2.1±0.3 nM（平均值±标准误，n = 3）的解离常数（KD）和58,400±1100个位点/细胞的最大结合量（Bmax）与SK-N-MC细胞中表达的Y1亚型神经肽Y受体可逆性结合。非特异性结合不超过KD时总放射性结合量的30%。在竞争实验中，[3H]BIBP3226被神经肽Y及其肽类似物浓度依赖性地取代，其亲和力模式为神经肽Y = [亮氨酸31，脯氨酸34]神经肽Y >> 神经肽Y-(18-36)。这种效价顺序与[3H]BIBP3226与Y1亚型神经肽Y受体的相互作用一致。因此，[3H]BIBP3226可作为研究神经肽Y Y1受体的选择性配体。