Purification, molecular cloning, and functional expression of the cholecystokinin receptor from rat pancreas
Stephen A. Wank,Richard N. Harkins,Robert T. Jensen,Hagit Shapira,A de Weerth,Timothy Slattery +5 more
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The purification of the CCKA receptor to homogeneity from rat pancreas is reported here by using ion-exchange and multiple affinity chromatographic separations to allow partial peptide sequencing after chemical/enzymatic cleavage and synthesis of degenerate oligonucleotide primers.Abstract:
The cholecystokinin (CCK) family of peptides and their receptors are widely distributed throughout the gastrointestinal and central nervous systems where they regulate secretion, motility, growth, anxiety, and satiety. The CCK receptors can be subdivided into at least two subtypes, CCKA and CCKB on the basis of pharmacological studies. We report here the purification of the CCKA receptor to homogeneity from rat pancreas by using ion-exchange and multiple affinity chromatographic separations. This allowed partial peptide sequencing after chemical/enzymatic cleavage and synthesis of degenerate oligonucleotide primers. These primers were used for initial cloning of the cDNA from rat pancreas by PCR. The predicted protein sequence of the cDNA clone contained the five partial peptide sequences obtained from the purified protein. Seven putative transmembrane domains suggest its membership in the guanine nucleotide-binding regulatory protein-coupled receptor superfamily. In vitro transcripts of the cDNA clone were functionally expressed in Xenopus oocytes and displayed the expected agonist and antagonist specificity.read more
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Free fatty acids regulate insulin secretion from pancreatic beta cells through GPR40.
Yasuaki Itoh,Yuji Kawamata,Masataka Harada,Makoto Kobayashi,Ryo Fujii,Shoji Fukusumi,Kazuhiro Ogi,Masaki Hosoya,Yasuhiro Tanaka,Hiroshi Uejima,Hideyuki Tanaka,Minoru Maruyama,Rie Satoh,Shoichi Okubo,Hideki Kizawa,Hidetoshi Komatsu,Fumika Matsumura,Yuko Noguchi,Tokuyuki Shinohara,Shuji Hinuma,Yukio Fujisawa,Masahiko Fujino +21 more
TL;DR: It is shown that a G-protein-coupled receptor, GPR40, which is abundantly expressed in the pancreas, functions as a receptor for long-chain FFAs, and that long- Chain FFAs amplify glucose-stimulated insulin secretion from pancreatic β cells by activating GPR 40.
Journal ArticleDOI
Peptidomimetics for Receptor Ligands—Discovery, Development, and Medical Perspectives
TL;DR: Peptidomimetics, compounds that act as substitutes for peptides in their interaction with receptors, have been synthesized and show higher metabolic stability, better bioavailability, and longer duration of action.
Journal ArticleDOI
Biological actions of cholecystokinin
TL;DR: New findings are presented on potent, nonpeptide CCK antagonists, selective for the two CCK receptor subtypes, which demonstrate that endogenous CCK has biologically important effects on physiology and behavior.
Journal ArticleDOI
Physiological regulation of G protein-linked signaling.
Andrew J. Morris,Craig C. Malbon +1 more
TL;DR: Physiological regulation of G protein-linked receptors allows for integration of signals that directly or indirectly effect the signaling from receptor-->G protein-->effector(s).
Journal ArticleDOI
Gastrointestinal hormones and food intake.
TL;DR: Although traditionally thought of as short-term, meal-related signals, enhanced, chronic hormone secretion and signaling resulting from gut reconstruction as seen with gastric bypass surgery most likely contributes to the superior efficacy of surgery as a treatment for obesity.
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Basic methods in molecular biology
TL;DR: General methods bacterial strains and cloning vectors enzymes that modify DNA and RNA in vitro amplification of DNA using the polymerase chain reaction (PCR) and the thermostable Taq DNA polymerase, introduction DNA restriction fragment analysis and preparation.