Wimalawansa S J, Gunasekera R D, Zhang F
Department of Chemical Pathology, Royal Postgraduate Medical School, London, UK.
Peptides. 1993 Jul-Aug;14(4):691-9. doi: 10.1016/0196-9781(93)90099-3.
Intact calcitonin gene-related peptide (CGRP) receptors were solubilized from porcine neural membranes using sodium cholate: potassium buffer. The solubilized receptors were purified sequentially by hydrophobic interaction and ion-exchange chromatography followed by specific affinity chromatography. Using these procedures, we have isolated 2 nmol of highly purified active CGRP receptor to a homogeneity (5 x 10(8)-fold purification). The isolated receptors retained their specificity and the capacity to bind to 125I-CGRP, and showed no cross-reactivity with a number of other peptides, except with amylin having 46% amino acid sequence homology to h-CGRP. The solubilized receptors were adsorbed by WGA-agarose and concanavalin-A, suggesting a glycoprotein nature. SDS-PAGE, size-exclusion HPLC, and autoradiography confirmed that CGRP receptor is a monomeric membrane protein with M(r) 66 kDa.
