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. 2013 Oct 9;4:2529. doi: 10.1038/ncomms3529

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Copyright © 2013, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.

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C83-FLAG and C99-FLAG substrates were incubated with a CHAPSO-solubilized CHO microsomal fraction to evaluate the substrate preference of γ-secretase (a). C83-FLAG was efficiently cleaved to produce AICD–FLAG compared with inefficient cleavage of C99-FLAG. Dashed lines indicate ε-cleavage sites. V1711 Notch-FLAG containing a native N terminus is a shorter substrate and was efficiently cleaved to produce NICD–FLAG compared with inefficient cleavage of ΔE Notch-FLAG (b). The dashed line indicates the S3 cleavage site. M664 APLP2-FLAG and R678 APLP2-FLAG are analogous to β- and α-cleaved substrates, respectively (c). R678 APLP2-FLAG possesses a short ectodomain and was efficiently cleaved by γ-secretase to produce AP2-ICD-FLAG compared with inefficient cleavage of M664 APLP2-FLAG. Dashed lines indicate cleavage sites.