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All known biological effects of CCK peptides reside in the conserved C-terminal heptapeptide sequence (Figure 1). Modification of this sequence grossly reduces or abolishes receptor binding and biological effects (13–15). The N-terminal extensions of the common C-terminus increase the biological potency and the specificity for receptor binding.
The release from neurons has been examined directly in brain slices and synaptosomes (56, 57). Potassium-induced depolarization caused a calcium-dependent release of CCK-8. Similarly, depolarization releases CCK peptides from the hypothalamic dopamine neurons that innervate the intermediate lobe of the pituitary (58). Cholecystokinin (CCK) is member of a family of regulatory peptides with a remarkably well preserved C-terminal sequence (1–3). The family also includes frog skin peptides (caerulein and phyllocaerulein) and the protochordean neuropeptide cionin, but in mammals, CCK and gastrin are the only family members (Figure 1). All relevant data were included in the manuscript or Supplementary information.
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- It is less specific than the CCK1 receptor and binds also non-sulfated CCK, gastrins, and C-terminal fragments such as CCK-5.
- It is worth noting that CCK1R binds and responds to sulfated CCK with a 500- to 1000-fold higher potency than non-sulfated CCK, whereas CCK2R discriminates poorly between sulfated and non-sulfated peptides45.
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Conceived and supervised the project and wrote the manuscript with inputs from all co-authors. CCK1R and CCK2R Gs-mediated Gs-cAMP accumulation assays were performed using HEK293T cells (ATCCCRL-11268) transiently expressing human CCK1R and the cAMP biosensor GloSensor-22F (Promega). Cells were seeded (4 × 103 cells, 40 μL/well) into 384-well culture plates and incubated for 24 h at 37 °C in 5% CO2. Next day, the culture medium was removed and the equilibration medium containing 4% (v/v) dilution of the GloSensor™ cAMP reagent stock solution was added to each well.
D The effect of Q204A mutation in CCK2R on CCK-8-induced Gαq–Gγ dissociation. Progressive jackpots pool a portion of bets across games or networks and grow until a lucky player hits the winning combination. Navigation is relatively straightforward, and loading speeds appear decent. There are numerous variants designed for international and local play.
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