Deposition of the amyloid-beta protein (An) is an early and invariant event in the pathogenesis of Alzheimer's disease (AD), and the two proteases that cut Abeta from the amyloid precursor protein (APP), beta- and y-secretases, are considered important therapeutic targets. y-Secretase catalyzes an unusual proteolysis within the single transmembrane region of APP, and this cleavage requires the multi-transmembrane presenilins. More than 60 different missense mutations in presenilins cause AD, and these mutations alter y-secretase activity to increase levels of a highly fibrillogenic 42-residue variant of Abeta (Abeta42). Inhibitor profiling, molecular modeling, and mutagenesis studies reveal that y-secretase has properties of an aspartyl protease and may catalyze an unusual intramembranous proteolysis. Moreover, two conserved transmembrane aspartates in presenilins are required for y-secretase activity, and transition-state analogue inhibitors of y-secretase bind directly and specifically to presenilins. Taken together, these results strongly suggest that presenilins themselves are y-secretases, novel intramembrane-cleaving aspartyl proteases. The objectives of the present proposal are to understand the topography of the y-secretase active site, to identify presenilin residues that are in or near the active site, and to develop compounds for studying the normal role of y-secretase in vivo and its potential as a therapeutic target for AD. The steric limits of y-secretase pockets S3 through S4' will be probed by systematically varying the size of the corresponding substituents in transition-state analogue inhibitors. The discovery of potent compounds would allow the development of affinity matrices for the purification of this protease and identification of associated cofactors. Specific regions within presenilins that interact with such inhibitors (i.e., parts in or near the active site) will be identified by photoaffinity labeling. The specific labeling of transmembrane residues of presenilins would provide compelling evidence for an intramembranous active site. Finally, nonpeptide analogues of these transition-state mimics will be developed using combinatorial chemistry as part of a strategy to identify metabolically stable compounds that block y-secretase activity in whole organisms.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS041355-03
Application #
6639747
Study Section
Bio-Organic and Natural Products Chemistry Study Section (BNP)
Program Officer
Murphy, Diane
Project Start
2001-04-01
Project End
2006-03-31
Budget Start
2003-04-01
Budget End
2004-03-31
Support Year
3
Fiscal Year
2003
Total Cost
$292,803
Indirect Cost
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
Wolfe, Michael S (2013) Toward the structure of presenilin/?-secretase and presenilin homologs. Biochim Biophys Acta 1828:2886-97
Wolfe, Michael S (2012) Processive proteolysis by ?-secretase and the mechanism of Alzheimer's disease. Biol Chem 393:899-905
Wolfe, Michael S (2012) ýý-Secretase inhibitors and modulators for Alzheimer's disease. J Neurochem 120 Suppl 1:89-98
Wolfe, Michael S (2012) ýý-Secretase as a target for Alzheimer's disease. Adv Pharmacol 64:127-53
Augelli-Szafran, C E; Wei, H-X; Lu, D et al. (2010) Discovery of notch-sparing gamma-secretase inhibitors. Curr Alzheimer Res 7:207-9
Wolfe, Michael S (2010) Structure, mechanism and inhibition of gamma-secretase and presenilin-like proteases. Biol Chem 391:839-47
Wolfe, Michael S (2009) gamma-Secretase in biology and medicine. Semin Cell Dev Biol 20:219-24
Wolfe, Michael S (2009) Intramembrane-cleaving proteases. J Biol Chem 284:13969-73
Wolfe, Michael S (2009) Intramembrane proteolysis. Chem Rev 109:1599-612
Wolfe, Michael S (2008) Inhibition and modulation of gamma-secretase for Alzheimer's disease. Neurotherapeutics 5:391-8

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