Blood brain barrier endothelial cells express candidate amyloid precursor protein-cleaving secretases.
Proteolytic cleavage of the amyloid precursor protein (A beta PP) results in the generation of the amyloidogenic fragment known as amyloid beta peptide (A beta). Deposition of A beta in the brain parenchyma and cerebrovasculature is a feature of Alzheimer's disease (AD). To date, the process whereby A beta is generated and deposited remains unclear. We have previously established that activated platelets from AD patients retain more A beta PP on their surface than control platelets. We report here that an endothelial cell-derived enzyme can cleave this surface platelet A beta PP. Human blood brain barrier endothelial cells from brains of AD patients were assayed for potential A beta PP-cleaving enzymes using synthetic peptide substrates encompassing the A beta N-terminus cleavage site. A protease activity capable of cleaving A beta PP on the surface of AD platelets was noted. The A beta PP cleavage is partially inhibited by EDTA, by ZincOV, as well as by a specific inhibitor of the Zn metalloprotease E.C.3.4.24.15. Furthermore, the protease is recognized by an antibody directed against it, using immunohistochemistry, Western blot analysis and flow cytometry. The protease is not secreted, but rather resides intracellularly as well as on the surface of the endothelial cells. The data suggest that E.C.3.4.24.15 synthesized by brain endothelial cells may process the platelet-derived A beta PP, yielding fragments which could contribute to cerebrovascular A beta deposits.
Simons
E R
ER
Department of Biochemistry, Boston University School of Medicine, MA, USA.
Marshall
D C
DC
Long
H J
HJ
Otto
K
K
Billingslea
A
A
Tibbles
H
H
Wells
J
J
Eisenhauer
P
P
Fine
R E
RE
Cribbs
D H
DH
Davies
T A
TA
Abraham
C R
CR
D000544:Alzheimer Disease / Q000201:enzymology*; D053829:Amyloid Precursor Protein Secretases; D016564:Amyloid beta-Protein Precursor / Q000378:metabolism*; D000918:Antibody Specificity; D016282:Aspartic Acid Endopeptidases; D001792:Blood Platelets / Q000378:metabolism; D001812:Blood-Brain Barrier; D004492:Edetic Acid / Q000494:pharmacology; D010450:Endopeptidases / Q000276:immunology / Q000302:isolation & purification*; D004730:Endothelium, Vascular / Q000201:enzymology*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D007150:Immunohistochemistry; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000276:immunology / Q000302:isolation & purification*; D009842:Oligopeptides / Q000378:metabolism
D016564:Amyloid beta-Protein Precursor; D006877:Hydroxamic Acids; D009842:Oligopeptides; C048727:zincov; D004492:Edetic Acid; D053829:Amyloid Precursor Protein Secretases; D010450:Endopeptidases; D016282:Aspartic Acid Endopeptidases; C509280:BACE1 protein, human; D008666:Metalloendopeptidases