鸽子小脑的一种红藻氨酸结合蛋白：通过单克隆抗体进行纯化和定位

A kainate binding protein in pigeon cerebellum: purification and localization by monoclonal antibody.

作者信息

Klein A U, Niederoest B, Winterhalter K H, Cuénod M, Streit P

机构信息

Brain Research Institute, University of Zürich, Switzerland.

出版信息

Neurosci Lett. 1988 Dec 19;95(1-3):359-64. doi: 10.1016/0304-3940(88)90685-4.

DOI:10.1016/0304-3940(88)90685-4

PMID:2852325

Abstract

A monoclonal antibody (mAb) was developed which precipitated kainate binding activity from detergent extract of pigeon cerebellum and was used to isolate a kainate binding protein (KBP) by immunoaffinity chromatography. It migrated at an apparent molecular weight (Mr) of 220,000 in gel filtration chromatography and at Mr = 50,000 in polyacrylamide gel electrophoresis performed under denaturing and reducing conditions. Moreover, the mAb produced an immunohistochemical staining pattern in the molecular and Purkinje cell layers which corresponded to the autoradiographic labeling pattern observed with tritiated kainate and were reported to be typical of the major kainate binding site in the pigeon cerebellum.

摘要

开发了一种单克隆抗体（mAb），它能从鸽子小脑的去污剂提取物中沉淀出海藻氨酸结合活性，并用于通过免疫亲和色谱法分离出海藻氨酸结合蛋白（KBP）。在凝胶过滤色谱中，它的表观分子量（Mr）为220,000，在变性和还原条件下进行的聚丙烯酰胺凝胶电泳中，Mr = 50,000。此外，该单克隆抗体在分子层和浦肯野细胞层产生的免疫组织化学染色模式，与用氚化海藻氨酸观察到的放射自显影标记模式相对应，据报道这是鸽子小脑主要海藻氨酸结合位点的典型特征。