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This smaller opening of the intracellular pocket for G protein is not suitable to engage with the Gs protein due to potential steric hindrance (Supplementary Fig. S5a). The α5 helix of Gαs protein is bulkier than that of other G proteins (Supplementary Fig. S5b) and thus it is believed that the engagement of Gs requires larger outward movement of TM6. Therefore, we hypothesized that the unique conformation of the intracellular half of the active CCK1R would lead to a non-canonical Gs-coupling mode that is elaborated below. This observation also supports the previous hypothesis that the smaller displacement of TM6 in the Gi/o-bound class-A GPCRs might preclude the binding of Gs49.
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- Apart from ligand selectivity, huge differences in downstream G protein signaling are present between CCK1R and CCK2R.
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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).