A homology model of inactive mouse FZD5 (UniProt ID: Q9EQD0) was generated using a crystal structure of SMO as template (PDB ID: 5V56) (28). To obtain a model of FZD5 in an active conformation, TM6 (residues 445 to 467) was modeled on the basis of the crystal structure of the β2AR in complex with Gs (PDB ID: 3SN6) (20). Because of the lack of suitable template, the N- and C-terminal regions (amino acids 1 to 228 and 541 to 585) were excluded from the models. Initial sequence alignments were generated using ClustalX2 (61), and homology models were constructed using Modeller 9.16 (62). A set of 100 homology models was generated for both the active and inactive conformations of FZD5. A representative model was selected among the 10 structures with the best discrete optimized protein energy (DOPE) scores (63) based on visual inspection.

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