Molecular cloning and functional characterization of a human secretin receptor.
Secretin is a gastrointestinal hormone responsible for the regulation of bicarbonate, potassium ion and enzyme secretion from the pancreas. A cDNA encoding the human secretin receptor was isolated from a human pancreatic adenocarcinoma cell-line cDNA library using polymerase chain reaction and library screening techniques. The cDNA isolated is 1717 bp in length encoding a 440 amino acid long polypeptide. Computer analysis of the receptor indicated that it is a member of the glucagon-VIP-secretin receptor family and is a G-protein coupled receptor containing seven hydrophobic transmembrane domains. The receptor was subsequently expressed in COS-7 cells and was able to bind specifically to human secretin with high affinity as indicated by the competitive displacement assay. The human secretin receptor was found to be functionally coupled to the stimulation of adenylyl cyclase resulting in the accumulation of intracellular cAMP in a dose-dependent manner. By Northern blot analysis, a 1.8 Kb mRNA was detected in human pancreas and intestine, while weak hybridization signals were detected in human colon, kidney and lung. Functional characterization of this receptor should enhance our understanding of the physiology and pathophysiology of human secretin, its structure-function, receptor interaction and receptor tissue distribution.
Chow
B K
BK
Department of Zoology, University of Hong Kong.
D000230:Adenocarcinoma; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D015152:Blotting, Northern; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D003106:Colon / Q000378:metabolism; D016384:Consensus Sequence; D017931:DNA Primers; D005260:Female; D015870:Gene Expression; D015723:Gene Library; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D007668:Kidney / Q000378:metabolism; D007700:Kinetics; D008168:Lung / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010179:Pancreas / Q000378:metabolism; D010190:Pancreatic Neoplasms; D016133:Polymerase Chain Reaction; D017433:Protein Structure, Secondary; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D051381:Rats; D043562:Receptors, G-Protein-Coupled; D011964:Receptors, Gastrointestinal Hormone / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D012633:Secretin / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014162:Transfection; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D012333:RNA, Messenger; D043562:Receptors, G-Protein-Coupled; D011964:Receptors, Gastrointestinal Hormone; D011994:Recombinant Proteins; C029937:secretin receptor; D012633:Secretin