Naldini L, Cirillo D, Moody T W, Comoglio P M, Schlessinger J, Kris R
Rorer Biotechnology, Inc., King of Prussia, Pennsylvania 19406.
Biochemistry. 1990 May 29;29(21):5153-60. doi: 10.1021/bi00473a022.
The receptor for the neuropeptide gastrin-releasing peptide, the mammalian homologue of bombesin, was solubilized from rat brain and Swiss 3T3 cells by using the zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid (CHAPS) and the cholesteryl hemisuccinate ester (CHS). Only the combination of the detergent CHAPS and the cholesteryl ester CHS in a glycerol-containing buffer satisfactorily preserved the binding activity upon solubilization. Specific binding activity was only solubilized from cell lines and tissue preparations known to express the GRP receptor. The dissociation constant (Kd) for the receptor solubilized from rat brain and Swiss 3T3 cells was 0.6 nM, similar to the value of 0.8 nM calculated for the membrane-bound receptor. Binding was saturable and reached equilibrium after approximately 2 h at 4 degrees C. The identity of the solubilized receptor with the membrane-bound one was further confirmed by the concordance of the relative binding affinities of various established bombesin analogues.
神经肽胃泌素释放肽（蛙皮素的哺乳动物同源物）的受体，通过使用两性离子去污剂3-[(3-胆酰胺丙基)二甲基铵]-1-丙烷磺酸（CHAPS）和胆固醇半琥珀酸酯（CHS），从大鼠脑和瑞士3T3细胞中溶解出来。只有在含甘油的缓冲液中，去污剂CHAPS和胆固醇酯CHS的组合在溶解时才能令人满意地保留结合活性。特异性结合活性仅从已知表达GRP受体的细胞系和组织制剂中溶解出来。从大鼠脑和瑞士3T3细胞中溶解出来的受体的解离常数（Kd）为0.6 nM，与为膜结合受体计算的0.8 nM值相似。结合是可饱和的，在4℃下约2小时后达到平衡。各种已确立的蛙皮素类似物的相对结合亲和力的一致性进一步证实了溶解的受体与膜结合受体的同一性。
