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- 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.
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Cryo-EM grid preparation and data collection
The concentration increases within 20 min to 3–5 pmol/l during meal stimulation, and then declines gradually only to reach a second peak after 1.5–2 hours. In comparison with most other pancreatic and gastrointestinal hormones (62), the concentrations of CCK in plasma are low. When food-induced CCK in plasma is mimicked by infusion of exogenous CCK, the same degree of gallbladder contraction and release of enzymes as seen during meals occurs (54, 62–64). Therefore, the low circulating concentrations of CCK are sufficient to account for the gallbladder contraction and pancreatic enzyme secretion during meals. The major sites of CCK expression are as mentioned endocrine cells in the gut, the brain, and in peripheral nerves.
Protein concentration was determined by absorbance at 280 nm using a Nanodrop 2000 Spectrophotometer (Thermo Fisher Scientific). The sample-coated grids were blotted before plunge-freezing into liquid ethane and stored in liquid nitrogen for data collection. The microscope was operated at 300 kV accelerating voltage, at a nominal magnification of 29,000 in counting mode, corresponding to a pixel size of 1.014 Å. A Electrostatic potential map of the binding pocket in CCK1R for the sulfated tyrosine of CCK-8. B The effect of N98A, R197A, or R197M mutation in CCK1R on CCK-8-induced cAMP accumulation. C Electrostatic potential map of the binding pocket in CCK2R for the sulfated tyrosine of CCK-8.
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