Nakata Y, Hiraoka C, Segawa T
Department of Pharmacology, Hiroshima University School of Medicine, Japan.
Eur J Pharmacol. 1988 Jul 26;152(1-2):171-4. doi: 10.1016/0014-2999(88)90851-5.
Substance P (SP) binding sites in rat cerebral cortical membranes were specifically and covalently labelled by means of disuccinimidyl suberate (DSS), a bifunctional cross-linking reagent, to determine the apparent molecular weight of the SP binding site. Cross-linking of [3H]SP by 1 mM DSS to membranes revealed a specifically labelled single protein with an apparent molecular weight of 46,000, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel filtration analyses. This labelling was inhibited by non-radioactive SP and was not changed by the presence or absence of GTPrS suggesting that the label was bound to the binding site of SP receptors.
为了确定P物质(SP)结合位点的表观分子量,使用双功能交联剂辛二酸二琥珀酰亚胺酯(DSS)对大鼠大脑皮层膜中的SP结合位点进行特异性共价标记。通过1 mM DSS将[3H]SP与膜交联,经十二烷基硫酸钠聚丙烯酰胺凝胶电泳和凝胶过滤分析测定,显示出一个表观分子量为46,000的特异性标记单一蛋白质。这种标记被非放射性SP抑制,并且不受GTPγS存在与否的影响,这表明该标记与SP受体的结合位点结合。