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From Figures 3 and 4, in particular, it is obvious that the newly formed Arg123-Glu40 salt bridge is due to the missense mutation L23B R (Figure 2), i.e., the substitutin of the leucine with a positively charged arginine at position 32 (instead of 23, Table 1 of supplementary file supps.pdf ) of semaglutides backbone (Figure 2)
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and resuspended in 20 mM HEPES pH 7.5, 100 mM NaCl, 10 mM MgCl 2 , 5 mM CaCl 2 , 2 mM KCl, 0.1 mM TCEP, supplemented with Protease Inhibitor Cocktail (TargetMol, 1 mL/100 mL suspension)
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