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P4‐299: A Computational Method to Predict Disease‐Specific Epitopes in Aβ, and its Application to Oligomer‐Selective Antibodies for Alzheimer’S Immunotherapy
Author(s) -
Plotkin Steven S.,
Peng Xubiao,
Silverman Judith M.,
Cashman Neil R.
Publication year - 2016
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2016.07.042
Subject(s) - oligomer , epitope , context (archaeology) , chemistry , molecular dynamics , proteostasis , neurodegeneration , conformational ensembles , protein folding , biophysics , dihedral angle , protein structure , computational biology , antibody , biology , biochemistry , computational chemistry , disease , molecule , immunology , medicine , paleontology , hydrogen bond , organic chemistry , pathology
in Alzheimer’s disease (AD) is truncated at the N-terminus. Among these, accumulation of pGlu-modified amyloid (pGlu-Ab3-40/42) has been shown to correlate with disease progression and tau pathology. Therapeutic strategies targeting pGlu-Ab, e.g. inhibitors of glutaminyl cyclase (QC) that catalyzes pGlu-formation, are currently in clinical assessment. However, it is still unclear, which enzyme(s) might be responsible for N-truncation of Ab and thus, generation of QC substrates. Aim of the study was to characterize the potential formation of the precursor(s) of pGlu-Ab3-40/42 by Meprin b in cell culture.Methods:We expressed different APP proteins (wt or familial Alzheimer’s mutations) in the cell lines HEK293 and CHO. The APP processing and production was assessed using Western Blot analysis and ELISAs detecting N-truncated or full length Ab. Maldi-TOF mass spectrometry was used to analyze cleavage of APP-derived peptides. Results:An in vitro analysis of cleavage of APP-related peptides suggested specificity of Meprin b for the b-site of APP. The primary cleavage products were Ab(1-x), Ab(2-x) and, to a lesser extent, Ab(3-x). In cell culture, co-expression of APP and Meprin b, but not of its isoenzyme Meprin a, resulted in production of N-truncated Ab peptides, primarily Ab(2-40/42). Thereby, Meprin b cleaved preferably APPwt and not APPswedish, which contrasts to the b-secretase BACE. The cleavage of APP resulted also in minor amounts of Ab(3-40/42), which was revealed by addition of human QC and application of highly sensitive ELISA detecting pGlu-Ab3-40/42. Finally, analysis of human brain samples revealed upregulation of Meprin b, but not Meprin a, in AD. Meprin b positive astrocytes are found in the vicinity of pGlu-Ab containing deposits in human tissue. Conclusions: The data support a BACE-independent processing of APPwt that may contribute to formation of N-truncated Ab. These truncated forms, in turn, might be prone to further post-translational modification. Thus, Meprin b might represent a potential upstream target to suppress formation of pGlu-Ab.