Biomedical subjects

Ci Chen

Publications and source records attributed to Ci Chen.

3 recordsLinked to original sources

PubMed2005

Basal receptor activation by locally produced glucagon-like peptide-1 contributes to maintaining beta-cell function.

Glucagon-like peptide 1 (GLP-1) is a physiological stimulus of pancreatic beta-cell function. This enteroendocrine hormone is produced by intestinal L cells, and is delivered via the bloodstream to GLP-1 receptors (GLP-1Rs) on pancreatic beta-cells. In addition, there is evidence that beta-cell GLP-1Rs maintain sustained basal activity even in the absence of intestinal peptide, an observation that has raised the question whether these receptors have some degree of ligand-independent function. Here, we provide an alternative explanation for basal receptor activity based on our finding that biologically relevant amounts of fully processed GLP-1 are locally generated by insulinoma cell lines, as well as by alpha-cells of isolated rat islets in primary culture. Presence of GLP-1 was established by immunocytochemistry, as well as by selective ELISAs and bioassays of cell supernatants. A GLP-1R antagonist significantly reduced insulin secretion/production in beta-TC-6 insulinoma cells and isolated rat islets, suggesting a functionally important loop between locally produced GLP-1 and its cognate receptor. Treatment with this antagonist also inhibited the growth of beta-TC-6 cells. These observations provide novel insight into the function of insulin-producing cell lines and native beta-cells during in vitro culture, and they support the idea that locally produced GLP-1 may play a role in intra-islet regulation.

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PubMed2004

Ligand function at constitutively active receptor mutants is affected by two distinct yet interacting mechanisms.

It has been proposed that mutations that induce constitutive activity in G-protein-coupled receptors (GPCRs) concomitantly enhance the ability of partial agonists to trigger second-messenger signaling. Using the cholecystokinin type 2 receptor (CCK-2R) as a model system, we have explored whether this association applies to a diverse set of activating mutations. Consistent with established principles, constitutively active CCK-2Rs resulting from amino acid substitutions within the third intracellular loop each systematically increased partial agonist activities versus corresponding wild-type values. In contrast, activating mutations within transmembrane domain segments near the extracellular loops led to an increase in efficacy of only a subset of compounds but decreased or did not change the function of others. When transmembrane domain amino acid substitutions were introduced in combination with intracellular amplifying mutations, observed changes in ligand activity were defined by the product of two discernible factors 1) systematic amplification caused by an equilibrium shift from the inactive to the active receptor conformation and 2) ligand-specific alterations in signaling, which probably result from mutation-induced changes in the putative binding pocket. These findings illustrate functional heterogeneity among GPCR mutants with ligand-independent signaling. A subgroup of activating mutations facilitates receptor isomerization to the active state and in parallel perturbs ligand receptor interactions. These mutants do not adhere to the previously proposed "hallmark criteria" of constitutive activity.

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PubMed2003

Identification of a series of CCK-2 receptor nonpeptide agonists: sensitivity to stereochemistry and a receptor point mutation.

The search for small-molecule drugs that act at peptide hormone receptors has resulted in the identification of a wide variety of antagonists. In contrast, the discovery of nonpeptide agonists has been far more elusive. We have used a constitutively active mutant of the cholecystokinin 2 receptor (CCK-2R) as a sensitive screen to detect ligand activity. Functional assessment of structural analogs of the prototype CCK-2R antagonist, L-365,260 [3R-N- (2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-N'-(3-methylphenyl)urea], resulted in the identification of a series of agonists. Each of the active molecules is an S enantiomer, whereas the corresponding R stereoisomers have little or no activity. Further in vitro and in vivo assessment at the wild-type receptor indicated that efficacy of the two most active ligands approached that of the endogenous hormone. The function of selected R and S enantiomers was differentially sensitive to a point mutation, N353L, within the putative CCK-2R ligand pocket. The results of this study highlight the potential of constitutively active receptors as drug screening tools and the interdependence of ligand stereochemistry and receptor conformation in defining drug efficacy.

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