CCK QUANTIFICATION AND RECEPTOR CHARACTERIZATION

Project: Research project

Project Details

Description

Our goal is to better understand the role of cholecystokinin (CCK) in
health and disease. This gut and brain hormone exists in multiple
molecular forms, ranging in size from 4 to 58 amino acids, with functions
including regulation of pancreatic secretion, gallbladder contraction,
gastrointestinal motility, trophic effects, and satiety. Our approach is 3
fold: a) characterization and quantification of CCK peptides, b)
functional and biochemical characterization of CCK receptors, and c)
description of the molecular events associated with CCK-receptor
interaction, including receptor activation and initiation of the effector
cascade. The peptide-related components include: 1) the chemical and mass
spectrometric elucidation of the primary structure of large human CCK
peptides, 2) the establishment and validation of complementary approaches
to the quantification of human CCK peptides (RIA, radioreceptor assay, and
bioassay), and 3) the experimental (fluorescence, UV, and CD spectroscopy
and NMR) and theoretical approaches to the determination of the
conformation of the receptor-binding region of CCK. The receptor-related
components include a major commitment to the development of new
methodology, with initial application to the pancreatic acinar cell
receptor; subsequent application will be to gastrointestinal smooth muscle,
studying preparations of fresh and cultured, healthy and neoplastic cells
and their membranes. We plan to develop a series of new probes for the
CCK-binding region of the receptor; these will include 1) short CCK
analogues with free amino groups available for chemical crosslinking, 2)
analogues with incorporated aryl nitrene and carbene precursors for
photoaffinity labeling, 3) anti-idiotypic antibodies, and 4)
naturally-occurring auto-antibodies. We also plan to generate a series of
immunologic probes for this receptor and its associated molecules, and to
solubilize, purify, and reconstitute these in a functional state. These
studies should further our understanding of the mechanisms whereby this
hormone interacts with its receptor to activate it and initiate the
effector cascade within target cells. This may help in the understanding
of potential aberrations of CCK-receptor interaction which may occur in
disease stages, and may have more general basic cellular and molecular
implications for the important process of message transduction across a
membrane.
StatusFinished
Effective start/end date8/1/833/31/16

ASJC

  • Medicine(all)