Metalloendopeptidase EC 3.4.24.15 is necessary for Alzheimer's amyloid-beta peptide degradation.
We have investigated the functional relationship between metalloendopeptidase EC 3.4.24.15 (MP24.15) and the amyloid precursor protein involved in Alzheimer's disease (AD) and discovered that the enzyme promotes Abeta degradation. We show here that conditioned medium (CM) of MP24.15 antisense-transfected SKNMC neuroblastoma has significantly higher levels of Abeta. Furthermore, synthetic-Abeta degradation was increased or decreased following incubation with CM of sense or antisense-transfected cells, respectively. Soluble Abeta1-42 was degraded more slowly than soluble Abeta1-40, while aggregated Abeta1-42 showed almost no degradation. Pretreatment of CM with serine proteinase inhibitors 4-(2-aminoethyl)benzenesulfonyl fluoride and diisopropyl fluorophosphate completely inhibited Abeta degradation. Additionally, alpha1-antichymotrypsin (ACT), a serpin family inhibitor tightly associated with plaques and elevated in AD brain, blocked up to 60% of Abeta degradation. Interestingly, incubation of CM of MP24. 15-overexpressing cells with ACT formed an SDS-resistant ACT complex, suggesting an ACT-serine proteinase interaction. Recombinant MP24. 15 alone did not degrade Abeta. 14C-Diisopropyl fluorophosphate-radiolabeled CM from MP24.15-overexpressing cells contained increased levels of several active serine proteinases, suggesting that MP24.15 activates one or more Abeta-degrading serine proteases. Thus, ACT may cause Abeta accumulation by inhibiting an Abeta-degrading enzyme or by direct binding to Abeta, rendering it degradation-resistant. Identification of the Abeta-degrading enzyme and MP24.15's role in its activation is underway. Pharmacological modulation of either enzyme may provide a means of regulating Abeta in the brain.
Yamin
R
R
Department of Biochemistry and Medicine, Boston University School of Medicine, Boston, Massachusetts 02118, USA.
Malgeri
E G
EG
Sloane
J A
JA
McGraw
W T
WT
Abraham
C R
CR
D000544:Alzheimer Disease / Q000378:metabolism*; D016564:Amyloid beta-Protein Precursor / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008666:Metalloendopeptidases / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured
D016564:Amyloid beta-Protein Precursor; D015842:Serine Proteinase Inhibitors; D008666:Metalloendopeptidases; C022510:thimet oligopeptidase