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. 2015 Jun 19;5:11322. doi: 10.1038/srep11322

Figure 3. TAT-APPsweBBP strongly binds to and is cleaved by BACE1.

Figure 3

(A) Biotin-labeled TAT-APPsweBBP was incubated with or without recombinant BACE1 protein and examined by WB. Full length TAT-APPsweBBP and β-cleaved TAT-APPsweBBP were clearly detected. (B) Immunoprecipitation (IP) with anti-BACE1 antibody and subsequent WB with anti-biotin antibody and (C) IP with anti-biotin antibody and then WB with anti-BACE1 antibody both reveal that TAT-APPsweBBP, but not TAT peptide, binds to BACE1. (D) To further confirm TAT-APPsweBPP can bind to BACE1, human wild-type APP expressing Chinese hamster ovary (CHO/APPwt) cells were incubated with biotin-labeled TAT-APPsweBBP or biotin-labeled APPsweBBP at 20 μM for 30 min, and then stained with anti-BACE1 and anti-biotin antibodies. TAT-APPsweBPP showed a strong association with BACE1 in both cytoplasmic membrane and intracellular compartments. (E) In addition, these treated cells were further analyzed by IP with anti-biotin and subsequent WB with anti-BACE1 antibodies, with results showing that TAT-APPsweBBP strongly and specifically binds to BACE1.