Solubilization of the bombesin receptor from Swiss 3T3 cell membranes

FEBS Letters

Coffer, Arnold¹  Sinnett-Smith, James¹  Fabregat, Isabel¹  Rozengurt, Enrique¹ 

[1] Imperial Cancer Research Fund, PO Box 123, Lincoln's Inn Fields, London WC2A 3PX, UK

关键词: Signal transduction;    Growth control;    Neuropeptide;   

DOI  :  10.1016/0014-5793(90)80710-Z

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Bombesin and structurally related peptides including gastrin releasing peptide (GRP) are potent mitogens for Swiss 3T3 cells. Here we attempted to solubilize bombesin receptors under conditions in which the ligand (¹²⁵I-labelled GRP) was prebound to the receptor prior to detergent extraction. We found that ¹²⁵I-GRP-receptor complexes were solubilized from Swiss 3T3 cell membranes by using the detergents taurodeoxycholate or deoxycholate. These detergents promoted ligand-receptor solubilization in a dose-dependent manner. In contrast, a variety of other detergents including Triton X-100, octylglycoside, CHAPS, digitonin, cholic acid and n-dodecyl-β-D-maltoside, were much less effective. Addition of guanosine 5'-[γ-thio]triphosphate (GTPγS) to ligand-receptor complexes isolated by gel nitration enhanced the rate of ligand dissociation in a concentration-dependent and nucleotide-specific manner. Our results demonstrate for the first time the successful solubilization of ¹²⁵ I-GRP-receptor complexes from Swiss 3T3 cell membranes and provide evidence for the physical association between the ligand-receptor complex and a guanine nucleotide binding protein(s).

【 授权许可】

Unknown