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Cryo-EM grid preparation and data collection
According to the two CCK-8-bound structures, the interfaces between CCK-8 and CCKRs and the overall shape of the orthostatic pockets are conserved, consistent with previous prediction45. However, the sub-pocket for the TYS of CCK-8 is more positively charged in CCK1R attributed to N982.61, and R197ECL2 contributes to a beneficial charge–charge interaction with TYS of CCK-8 (Fig. 3a). Site-directed mutations R197ECL2M/A or N982.61A in CCK1R almost abolished CCK-8-triggered activation, implying the critical role of ECL2 in sulfated endogenous ligand recognition (Fig. 3b). The substitution of Q204ECL2A, the closest polar residue in CCK2R compared to CCK-8, does not affect CCK-8-induced Gq signaling of CCK2R measured by NanoBiT assay (Fig. 3d).
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Gut Motility
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- The picture has varied (12) due to species differences and because the molecular pattern along the gut varies (59, 60).
- CCK peptides also stimulate calcitonin, insulin, and glucagon secretion, and they may act as natriuretic peptides in the kidneys.
- CCK1R TM6 locates between the positions of β2AR TM6 in the inactive and active states (Fig. 5c).
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Meanwhile, various CCKR-targeted drugs have been developed, yet most of them were ultimately removed from the market due to questionable efficacy, lack of target specificity, or severe adverse effects28,29. This highlights the need for investigation of the structure of CCK1R and CCK2R to understand agonist binding, receptor activation and thus promote structure-based drug discovery. The biochemical concept of CCK as a single hormonal peptide from the small intestine has also changed considerably. Now CCK is known to be synthetized and released in multiple molecular forms. And the CCK gene is expressed at peptide level in a cell-specific manner in neurons, endocrine cells, and epithelial cells outside the gastrointestinal tract (Table 1).
