Properties of a recombinant human secretin receptor: a comparison with the rat and rabbit receptors.
A secretin receptor was cloned from a commercial human pancreatic complementary DNA (cDNA) bank. The amino acid sequence deduced from the nucleotide sequence differed slightly from the three different sequences previously published, suggesting a genetic polymorphism of the human receptor. The binding properties of the receptor were evaluated by testing natural secretin, related peptides, and synthetic analogs or fragments on membranes of Chinese hamster ovary (CHO) cells expressing the receptor after transfection. The second-messenger coupling was evaluated by adenylate cyclase measurement. The human secretin receptor was compared with the rat and the rabbit receptors. In the three animals species, rat and human secretin were equipotent; rabbit secretin was equipotent on human and rabbit secretin receptors and less potent on the rat receptor. Similar data were obtained for the [Arg16]-secretin analog. Deletion of histidine 1 and replacement of aspartate 3 reduced the affinity of the peptides for the three receptors; however, the reduction was more pronounced on rat than on human and rabbit secretin receptors. Finally, the low affinity of the rat and human receptors for vasoactive intestinal peptide (VIP) was identical; the rabbit receptor, however, had a 20-fold higher affinity. Thus the human secretin receptor shows properties of both rat and rabbit receptors. Evaluation of the properties of chimeric receptors will be useful to fit the ligand on the receptors.
Di Paolo
E
E
Department of Biochemistry, Faculty of Medicine, Brussels, Belgium.
De Neef
P
P
Moguilevsky
N
N
Petry
H
H
Cnudde
J
J
Bollen
A
A
Waelbroeck
M
M
Robberecht
P
P
D000262:Adenylyl Cyclases / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells / Q000378:metabolism; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000032:analysis; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D010179:Pancreas; D011817:Rabbits; D051381:Rats; D043562:Receptors, G-Protein-Coupled; D011964:Receptors, Gastrointestinal Hormone / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins; D012633:Secretin / Q000235:genetics* / Q000378:metabolism; D013045:Species Specificity; D014162:Transfection; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism
D018076:DNA, Complementary; D043562:Receptors, G-Protein-Coupled; D011964:Receptors, Gastrointestinal Hormone; D011994:Recombinant Proteins; C029937:secretin receptor; D012633:Secretin; D014660:Vasoactive Intestinal Peptide; D000262:Adenylyl Cyclases