ABSTRACT
BACKGROUND: Amyloid-ß peptide ending at 42nd residue (Aß42) is believed as a pathogenic peptide for Alzheimer disease. Although γ-secretase is a responsible protease to generate Aß through a processive cleavage, the proteolytic mechanism of γ-secretase at molecular level is poorly understood. RESULTS: We found that the transmembrane domain (TMD) 1 of presenilin (PS) 1, a catalytic subunit for the γ-secretase, as a key modulatory domain for Aß42 production. Aß42-lowering and -raising γ-secretase modulators (GSMs) directly targeted TMD1 of PS1 and affected its structure. A point mutation in TMD1 caused an aberrant secretion of longer Aß species including Aß45 that are the precursor of Aß42. We further found that the helical surface of TMD1 is involved in the binding of Aß45/48 and that the binding was altered by GSMs as well as TMD1 mutation. CONCLUSIONS: Binding between PS1 TMD1 and longer Aß is critical for Aß42 production.
Subject(s)
Amyloid beta-Peptides/metabolism , Peptide Fragments/metabolism , Presenilin-1/metabolism , Amyloid beta-Peptides/chemistry , Animals , Humans , Mice , Mice, Inbred C57BL , Mice, Knockout , Protein Structure, Quaternary , TransfectionABSTRACT
Substrate-selective inhibition or modulation of the activity of γ-secretase, which is responsible for the generation of amyloid-ß peptides, might be an effective strategy for prevention and treatment of Alzheimer's disease. We have shown that helical ß-peptide foldamers are potent and specific inhibitors of γ-secretase. Here we report identification of target site of the foldamers by using a photoaffinity probe. The photoprobe directly and specifically labeled the N-terminal fragment of presenilin 1, in which the initial substrate docking site is predicted to be located. We also optimized the foldamer structure by preparing a variety of derivatives and obtained two highly potent foldamers by incorporation of a hydrophilic and neutral functional group into the parent structure. The class of side chain functional group and the position of incorporation were both important for γ-secretase-inhibitory activity. The substrate selectivity of the inhibitory activity was also quite sensitive to the class of side chain group incorporated.
Subject(s)
Amyloid Precursor Protein Secretases/antagonists & inhibitors , Amyloid Precursor Protein Secretases/chemistry , Oligopeptides/chemistry , Cyclopentanes/chemical synthesis , Cyclopentanes/chemistry , Cyclopentanes/pharmacology , HEK293 Cells , Humans , Hydrophobic and Hydrophilic Interactions , Molecular Mimicry , Oligopeptides/chemical synthesis , Oligopeptides/pharmacology , Proline/analogs & derivatives , Proline/chemical synthesis , Proline/chemistry , Proline/pharmacology , Protein Structure, Secondary , Stereoisomerism , Substrate SpecificityABSTRACT
Amyloid-ß peptide ending at the 42nd residue (Aß42) is implicated in the pathogenesis of Alzheimer's disease (AD). Small compounds that exhibit selective lowering effects on Aß42 production are termed γ-secretase modulators (GSMs) and are deemed as promising therapeutic agents against AD, although the molecular target as well as the mechanism of action remains controversial. Here, we show that a phenylpiperidine-type compound GSM-1 directly targets the transmembrane domain (TMD) 1 of presenilin 1 (PS1) by photoaffinity labelling experiments combined with limited digestion. Binding of GSM-1 affected the structure of the initial substrate binding and the catalytic sites of the γ-secretase, thereby decreasing production of Aß42, possibly by enhancing its conversion to Aß38. These data indicate an allosteric action of GSM-1 by directly binding to the TMD1 of PS1, pinpointing the target structure of the phenylpiperidine-type GSMs.
Subject(s)
Allosteric Regulation/drug effects , Amyloid Precursor Protein Secretases , Amyloid beta-Peptides/metabolism , Peptide Fragments/metabolism , Piperidines/pharmacology , Presenilin-1 , Protein Conformation/drug effects , Alzheimer Disease/physiopathology , Amyloid Precursor Protein Secretases/antagonists & inhibitors , Amyloid Precursor Protein Secretases/metabolism , Amyloid beta-Peptides/analysis , Animals , Binding Sites/drug effects , CHO Cells , Cricetinae , Cricetulus , HEK293 Cells , Humans , Mice , Mice, Inbred C57BL , Peptide Fragments/analysis , Piperidines/chemical synthesis , Presenilin-1/chemistry , Presenilin-1/metabolism , Protein Binding/drug effectsABSTRACT
Sphingosine kinase (SphK) 1 and 2 phosphorylate sphingosine to generate sphingosine-1-phosphate (S1P), a pluripotent lipophilic mediator implicated in a variety of cellular events. Here we show that the activity of ß-site APP cleaving enzyme-1 (BACE1), the rate-limiting enzyme for amyloid-ß peptide (Aß) production, is modulated by S1P in mouse neurons. Treatment by SphK inhibitor, RNA interference knockdown of SphK, or overexpression of S1P degrading enzymes decreased BACE1 activity, which reduced Aß production. S1P specifically bound to full-length BACE1 and increased its proteolytic activity, suggesting that cellular S1P directly modulates BACE1 activity. Notably, the relative activity of SphK2 was upregulated in the brains of patients with Alzheimer's disease. The unique modulatory effect of cellular S1P on BACE1 activity is a novel potential therapeutic target for Alzheimer's disease.
