Volume 272, Number 34, Issue of August 22, 1997 pp. 21201-21206
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

High Potency Antagonists of the Pancreatic Glucagon-like Peptide-1 Receptor

(Received for publication, November 19, 1996, and in revised form, April 17, 1997)

Chahrzad Montrose-Rafizadeh , Huan Yang , Buel D. Rodgers , Alvie Beday , Louella A. Pritchette and John Eng ^¶

From the  Laboratory of Clinical Physiology, NIA, National Institutes of Health, Baltimore, Maryland 21224, the ^¶ Department of Medicine, Veterans Administration Medical Center, Bronx, New York 10468, and the  Mt. Sinai School of Medicine, New York, New York 10029

GLP-1-(7-36)-amide and exendin-4-(1-39) are glucagon-like peptide-1 (GLP-1) receptor agonists, whereas exendin-(9-39) is the only known antagonist. To analyze the transition from agonist to antagonist and to identify the amino acid residues involved in ligand activation of the GLP-1 receptor, we used exendin analogs with successive N-terminal truncations. Chinese hamster ovary cells stably transfected with the rat GLP-1 receptor were assayed for changes in intracellular cAMP caused by the test peptides in the absence or presence of half-maximal stimulatory doses of GLP-1. N-terminal truncation of a single amino acid reduced the agonist activity of the exendin peptide, whereas N-terminal truncation of 3-7 amino acids produced antagonists that were 4-10-fold more potent than exendin-(9-39). N-terminal truncation of GLP-1 by 2 amino acids resulted in weak agonist activity, but an 8-amino acid N-terminal truncation inactivated the peptide. Binding studies performed using ¹²⁵I-labeled GLP-1 confirmed that all bioactive peptides specifically displaced tracer with high potency. In a set of exendin/GLP-1 chimeric peptides, substitution of GLP-1 sequences into exendin-(3-39) produced loss of antagonist activity with conversion to a weak agonist. The results show that receptor binding and activation occur in separate domains of exendin, but they are more closely coupled in GLP-1.

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