Molecular structure analysis of the pituitary adenylate cyclase activating polypeptide type I receptor from pig brain

Photoaffinity Labeling Analysis of the Interaction of Pituitary Adenylate‐Cyclase‐Activating Polypeptide (PACAP) with the PACAP Type I Receptor

“…A protein band of M , 66000 was observed. The molecular mass of this labeled protein is identical to the mass of the porcine PACAP type I receptor determined by cross-linking with disuccinimidyl suberate, as previously described (Cao et al, 1994). Complete inhibition of the labeled products with 500-fold excess of PACAP-(1 -27)-peptide demonstrated the specificity of the photoaffinity labeling.…”

“…CF,CO,H/ phenol/water/triisopropylsilane (Sole and Barany, 1992 Radioiodination of PACAP-(1-27)-peptide and its photoreactive analogues. PACAP-( 1 -27)-peptide was radioiodinated by iodogen and was purified by HPLC as previously described (Cao et al, 1994). The photoreactive analogues were also labeled by the iodogen technique.…”

“…Crude membranes from transfected COS cells and pig brain were prepared as previously described (Cao et al, 1994(Cao et al, , 1995. The designated membrane fragments were frozen in aliquots and stored at -70°C.…”

“…Some structural requirements for the occupancy of the PACAP type I receptors have been reported and led to the development of receptor antagonists Vandermeers et al, 1992;Bitar et al, 1994). By using chemical modification and deletion mutagenesis, we have previously demonstrated that the N-terminal fragment of the PACAP type I receptor contains a major binding site for its ligand (Cao et al, 1994(Cao et al, , 1995. In the present study, we used photoaffinity labeling, which provides a method to directly identify contact regions of a receptor and its ligand.…”

Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) 38 and PACAP27 Activate Common and Distinct Intracellular Signaling Pathways to Stimulate Growth Hormone Secretion from Porcine Somatotropes1

“…Initial studies suggested that the response of somatotropes to PACAP was mediated by VPAC 2 or VPAC 1 , but not PAC 1 (2,23,27,28). Although the type of PVRs expressed in porcine pituitary has not yet been determined, a type I PACAP-binding site, analogous to the rat PAC 1 , has been isolated from pig brain (49,50). Although the type of PVRs expressed in porcine pituitary has not yet been determined, a type I PACAP-binding site, analogous to the rat PAC 1 , has been isolated from pig brain (49,50).…”