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神经母细胞瘤和杂交细胞系中δ-阿片受体33 kDa蛋白亚基的光亲和标记

Photoaffinity labeling of a 33 kDa protein subunit of the delta-opioid receptor in neuroblastoma and hybrid cell lines.

作者信息

Icard-Liepkalns C, Bochet P

机构信息

Laboratoire de Physiologie Nerveuse, CNRS, Gif-sur-Yvette, France.

出版信息

FEBS Lett. 1988 Jun 6;233(1):167-72. doi: 10.1016/0014-5793(88)81377-2.

Abstract

Tritiated DTLET (Tyr-D-Thr-Gly-Phe-Leu-Thr) binds with high affinity, specificity and saturability to neuroblastoma N18TG2 and hybrid neuroblastoma x glioma NG108-15 and NG108-5 intact cells. The delta-opioid receptor density in cells cultured in chemically defined medium was increased about 2 times compared to that in cells cultured in 10% fetal calf serum. A major and a minor protein species covalently and specifically bound to [125I]azido-DTLET (Tyr-D-Thr-Gly-pN3Phe-Leu-Thr), photoactivatable ligand, migrated on SDS-gel electrophoresis with Mr values near 33,000 and 58,000, respectively.

摘要

氚标记的DTLET(酪氨酸-D-苏氨酸-甘氨酸-苯丙氨酸-亮氨酸-苏氨酸)以高亲和力、特异性和饱和性与神经母细胞瘤N18TG2以及杂交神经母细胞瘤x胶质瘤NG108 - 15和NG108 - 5完整细胞结合。与在10%胎牛血清中培养的细胞相比,在化学成分确定的培养基中培养的细胞中δ阿片受体密度增加了约2倍。一种主要和一种次要蛋白质物种与光可活化配体[125I]叠氮-DTLET(酪氨酸-D-苏氨酸-甘氨酸-pN3苯丙氨酸-亮氨酸-苏氨酸)共价且特异性结合,在SDS凝胶电泳上迁移,其分子量分别接近33,000和58,000。

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