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As an example of the structural electrostatic perspective, in 2021, a manually designed structural modification involving a Val27-Arg28 (Val33-Arg34 in PDB entry 4ZGM) exchange (V-R exchange) [43] was introduced into the backbone of semaglutide towards the enhancement of semaglutide-GLP-1R binding affinity (K d ) from 3.4 10 6 M at 37 C to 1.1 10 6 M at 37 C, where a newly established structure-stabilizing interfacial salt bridge was formed between Glu104 (A Glu 127 in PDB entry 4ZGM) of GLP-1R ECD and Arg27 (B Val 33 in PDB entry 4ZGM) of semaglutide, which is entirely due to the manually designed V-R exchange [43]