Apolipoprotein E4: A causative factor and therapeutic target in neuropathology, including Alzheimer’s disease

Mahley et al. 10.1073/pnas.0600549103.

Supporting Information

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Supporting Figure 5
Supporting Table 2
Supporting Figure 6

Fig. 5. ApoE fragmentation in the brains of NSE-apoE or GFAP-apoE mice and humans. C-terminal-truncated fragments of 29-30 kDa were found in the brains of transgenic mice expressing apoE4 in neurons (NSE-apoE4 mice) and in the brains of patients with Alzheimer’s disease (1-3). ApoE in brain lysates of NSE-apoE (A) or GFAP-apoE (B) mice or humans (C) were detected by Western blotting with antibodies against full-length apoE or the C-terminal 28 aa of apoE. Note that the apoE fragmentation occurs in NSE-apoE mouse brains (A), which is similar to that in human brains (C), but not in GFAP-apoE mouse brains (B). All of the fragments lacked the C-terminal 27 aa. A first cleavage site yielded fragments of 29-30 kDa lacking the C-terminal 27 aa. In brains of Alzheimer’s disease patients and NSE-apoE4 mice, fragments of 15–20 kDa were also observed. (D) Mutation of Arg-61 to Thr or Glu-255 to Ala demonstrates that domain interaction is responsible for the susceptibility of apoE4 to proteolysis (3). (Modified from ref. 3.)

1. Huang, Y., Liu, X. Q., Wyss-Coray, T., Brecht, W. J., Sanan, D. A. & Mahley, R. W. (2001) Proc. Natl. Acad. Sci. USA 98, 8838–8843.

2. Harris, F. M., Brecht, W. J., Xu, Q., Tesseur, I., Kekonius, L., Wyss-Coray, T., Fish, J. D., Masliah, E., Hopkins, P. C., Scearce-Levie, K., et al. (2003) Proc. Natl. Acad. Sci. USA 100, 10966–10971.

3. Brecht, W. J., Harris, F. M., Chang, S., Tesseur, I., Yu, G.-Q., Xu, Q., Fish, J. D., Wyss-Coray, T., Buttini, M., Mucke, L., et al. (2004) J. Neurosci. 24, 2527–2534.

Supporting Figure 6

Fig. 6. Enhanced apoE4-induced Ab production abolished by treatment with small molecules. Treatment of apoE3 with GIND-25 and GIND-105 had no effect on the apoE3-enhanced Ab production; however, these small molecules significantly reduced the apoE4-enhanced Ab production and converted apoE4 to a molecule functionally resembling apoE3. GIND-25, azocarmine G; GIND-105, 3-butyl-1-ethyl-5-[2-(3-sulfobutyl-benzo[1,3]oxazolin-2-ylidene)-ethylidene]-2-thioxo-imidazolidin-4-one potassium salt. (Image reproduced with permission from ref. 1.)

1. Ye, S., Huang, Y., Müllendorff, K., Dong, L., Giedt, G., Meng, E. C., Cohen, F. E., Kuntz, I. D., Weisgraber, K. H. & Mahley, R. W. (2005) Proc. Natl. Acad. Sci. USA 102, 18700–18705.

Table 2. Cell-penetrating peptides (positively charged domain)

HIV-1 Tat	YGRKKRRQRRR
HSV VP22	DAATATRGRSAASTPTERPRAPARSASRPRRPVE
ANTP	RQIKIWPQNRRMKWKK
NF-kB p50	RRKSDLETSEPKPFLYYPEIKDKEEVQRKRQK
ApoE 134–158	RVRLASHLRKLRKRLLRDADDLQKR