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The mutation of GLP-1 Phe 28 to Ala causes it to lose nearly all of its affinity (IC 50 : 0.27 nM→357 nM), and Ile 29 mutation decreases the affinity notably (IC 50 : 0.27 nM→25 nM), while Trp 31 , Leu 32 , and Val 33 mutations have little influence, showing that they may be merely subsidiary