Figure 1: AEP cleaves APP in vitro. | Nature Communications

Figure 1: AEP cleaves APP in vitro.

From: Delta-secretase cleaves amyloid precursor protein and regulates the pathogenesis in Alzheimer’s disease

Figure 1

(a) APP cleavage by AEP+/+ kidney lysates. APP was cleaved at pH 6.0 by AEP+/+ kidney lysates (left panels). The enzymatic activity of AEP was determined using AEP activity assay (right panel; mean±s.e.m. of three independent experiments). (b) Antibody titration assay. Kidney lysates were incubated with AEP-specific antibody for 5 min, and then incubated with GST-APP for 30 min. The processing of APP was determined using western blot. (c) Mutants of AEP C189 and N323 diminish APP cleavage. (d) Blocked of APP cleavage by Fmoc-Ala-Glu-Asn-Lys-NH2 (AENK) peptide (left panel). AEP activity was inhibited by AENK but not by Fmoc-Ala-Glu-Gln-Lys-NH2 (AEQK) (right panel; mean±s.e.m.; n=3). (e) Cleavage of endogenous APP by AEP. (f) Western blot showing the processing of purified GST-APP by recombinant AEP. The AEP-derived APP fragments were detected using anti-APP N-terminal antibody (left panel) and anti-APP C-terminal antibody (right panel). a.f.u., arbitrary fluorescence unit; DMSO, dimethylsulfoxide; MW, molecular weight; WB, western blot.

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