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Western blotting showed that the quantity of Aβ made by the L30F/T164A mutant was close to twice the quantity produced by the WT (Figure 6b). We utilized ELISA to quantify Aβ40 and also aβ42 output through the L30F/T164A mutant; we uncovered that the L30F/T164A mutation resulted in roughly twice as much Aβ40 in addition to aβ42, compared While using the quantities made by the WT (Figure 6c). The ratio of prolonged Aβ42 using aβ40 was similar amongst the WT and mutant (Figure 6d).
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The distance among the oxygen atom of Thr164 of Aph1 and the oxygen atom of Tyr466 of PS1 is predicted to become three.five Å, suggesting that a hydrogen bond (H...O length, approximately 1.eight Å) is formed in these side chains (Figure 7b). The T164A mutation may possibly remove the hydrogen bond with PS1 Y466, resulting in versatile movement from the carboxy terminus of PS1. We suggest that Aph1 mutations control protease website activity by means of conformational improvements in PS1.
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