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For Y2H assays, the plasmids of pGADT7 (empty vector) or pGADT7 derivatives were introduced into the yeast strain AH109 with plasmids of pGBKT7 (empty vector) or pGBKT7 derivatives

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Gln64 serves as the primary ligand for MDSA or EA binding, while Tyr562 forms a hydrogen bond with Gln64