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The particles were extracted and subjected to 3D classification on the complex, and the best-resolved class was selected. The resulting set of 299,092 particles was subjected to CTF refinement, Bayesian polishing and 3D auto refinement using the pixel size of 1.014 Å. The final map has an indicated global resolution of 3.2 Å at a Fourier shell correlation (FSC) of 0.143.
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- In the CCK2R–Gq complex, Gq undergoes an anticlockwise rotation relative to the receptor compared to Gs (except for EP4R–Gs) (Supplementary Fig. S5a, f).
Adrenal medullary cells produce small amounts of CCK, although amidated and with a low degree of sulfation (98). The picture has varied (12) due to species differences and because the molecular pattern along the gut varies (59, 60). In man, CCK-33 predominates in plasma, but CCK-58, -22, and -8 are also present (11, 61). Moreover, cerebral CCK neurons are more abundant than neurons of other neuropeptides (42, 45, 46). While most peptidergic neurons occur in subcortical regions, CCK is expressed in the highest concentrations in neocortical neurons (39, 42, 47). The perikarya of the cortical CCK nerves are distributed in layers II–VI, with the highest frequency in layers II and III (42, 48).
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In this paper, we reported four high-resolution cryo-EM structures of CCK1R and CCK2R signaling complexes. Sulfation of CCK and gastrin in the body is highly regulated by sulfotransferase (SULT) enzymes with tissue-specific expression patterns, e.g., SULT4 is predominantly expressed in the brain61. The sulfation level of CCK and gastrin is also correlated with sub-cellular and cellular distribution. Interestingly, the non-sulfated CCK is enriched in neuronal cell bodies, whereas the sulfated CCK predominantly distributes in the nerve terminals62. CCKRs differentially recognize the sulfated and non-sulfated ligands enabling subtle and precisely regulated responses to extracellular stimuli.