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Although the single-residue mutations of CCK1R to the corresponding residue in CCK2R (i.e., S149A ICL2 , N304K 6.26 , and N374H 8.47 ) caused little change to CCK1R-mediated G s signaling, surprisingly, the triple-mutation of CCK1R (S149A ICL2 /N304K 6.26 /N374H 8.47 ) completely abolished G s signaling while only slightly affecting G q signaling of CCK1R, indicating that the combination of three amino acids collectively contributes to the selectivity of CCKRs towards G s or G q for CCKR family, especially essential for the G s coupling of CCK1R (Fig