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When superimposing the cryo-EM structure of the Nedd4-2 C2 domain on available crystal structures of Nedd4-2 and Nedd4-1 isoform 4 C2 domains (PDB: 2NSQ and 3B7Y), we noted that the Ca 2+ -binding loops had a different conformation in the cryo-EM structure, possibly due to interactions with the HECT N-lobe (Supplementary Fig

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