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Next, to further distinguish the significance of the two metal-binding sites in calcium activation of Pc ncAAAD, we set out to destroy site A (SiteA-MT-Ca, Asp971, Glu972, Asp973, Glu638, and Glu637), site B (SiteB-MT-Ca, Glu824, Asp825, and Asp931), or both sites (SiteA+B-MT-Ca) with alanine substitutions of the aspartates and glutamates that engaged in the coordination of calcium (Fig
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