TY - JOUR
T1 - Characterization of Alzheimer's β-secretase protein BACE
T2 - A pepsin family member with unusual properties
AU - Haniu, Mitsuru
AU - Denis, Paul
AU - Young, Yunjen
AU - Mendiaz, Elizabeth A.
AU - Fuller, Janis
AU - Hui, John O.
AU - Bennett, Brian D.
AU - Kahn, Steven
AU - Ross, Sandra
AU - Burgess, Teresa
AU - Katta, Viswanatham
AU - Rogers, Gary
AU - Vassar, Robert
AU - Citron, Martin
PY - 2000/7/14
Y1 - 2000/7/14
N2 - The cerebral deposition of amyloid β-peptide is an early and critical feature of Alzheimer's disease. Amyloid β-peptide is released from the amyloid precursor protein by the sequential action of two proteases, β- secretase and γ-secretase, and these proteases are prime targets for therapeutic intervention. We have recently cloned a novel aspartic protease, BACE, with all the known properties of β-secretase. Here we demonstrate that BACE is an N-glycosylated integral membrane protein that undergoes constitutive N. terminal processing in the Golgi apparatus. We have used a secreted Fc fusion-form of BACE (BACE-IgG) that contains the entire ectodomain for a detailed analysis of post-translational modifications. This molecule starts at Glu46 and contains four N-glycosylation sites (Ash153, Ash172, Ash223, and Ash354). The six Cys residues in the ectodomain form three intramolecular disulfide linkages (Cys216-Cys420, Cys278-Cys443, and Cys330-Cys380). Despite the conservation of the active site residues and the 30-37% amino acid homology with known aspartic proteases, the disulfide motif is fundamentally different from that of other aspartic proteases. This difference may affect the substrate specificity of the enzyme. Taken together, both the presence of a transmembrane domain and the unusual disulfide bond structure lead us to conclude that BACE is an atypical pepsin family member.
AB - The cerebral deposition of amyloid β-peptide is an early and critical feature of Alzheimer's disease. Amyloid β-peptide is released from the amyloid precursor protein by the sequential action of two proteases, β- secretase and γ-secretase, and these proteases are prime targets for therapeutic intervention. We have recently cloned a novel aspartic protease, BACE, with all the known properties of β-secretase. Here we demonstrate that BACE is an N-glycosylated integral membrane protein that undergoes constitutive N. terminal processing in the Golgi apparatus. We have used a secreted Fc fusion-form of BACE (BACE-IgG) that contains the entire ectodomain for a detailed analysis of post-translational modifications. This molecule starts at Glu46 and contains four N-glycosylation sites (Ash153, Ash172, Ash223, and Ash354). The six Cys residues in the ectodomain form three intramolecular disulfide linkages (Cys216-Cys420, Cys278-Cys443, and Cys330-Cys380). Despite the conservation of the active site residues and the 30-37% amino acid homology with known aspartic proteases, the disulfide motif is fundamentally different from that of other aspartic proteases. This difference may affect the substrate specificity of the enzyme. Taken together, both the presence of a transmembrane domain and the unusual disulfide bond structure lead us to conclude that BACE is an atypical pepsin family member.
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U2 - 10.1074/jbc.M002095200
DO - 10.1074/jbc.M002095200
M3 - Article
C2 - 10887202
AN - SCOPUS:0034647752
SN - 0021-9258
VL - 275
SP - 21099
EP - 21106
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 28
ER -