gamma-Secretase is an unprecedented intramembrane-cleaving aspartyl protease necessary for life in multicellular organisms. It processes a broad and growing array of single-transmembrane proteins, including Notch, Delta/Jagged, CD44, cadherins and certain sodium channel subunits. gamma-Secretase was originally identified as the protease affecting the final scission of APP to release amyloid beta-protein, which is causally implicated in Alzheimer's disease (AD). Presenilin, mutations in which cause the most common and aggressive form of early-onset AD, contains two conserved intra-membrane aspartates that serve as the catalytic site. The gamma- secretase complex also contains 3 other integral membrane proteins: Nicastrin, Aph-1 and Pen-2. Because the protease complex contains 19 transmembrane domains, multiple glycosylation sites on Nicastrin, and are unstable in many relatively mild detergents, crystallographic approaches to its structure are difficult and remote. We recently purified the mammalian complex essentially to homogeneity and performed electron cryo- microscopy and single particle image analysis to obtain the first 3D map of the intramembrane protease. We now propose to build on this initial model by significantly improving the resolution of the structure. We will localize the catalytic site of the gamma-secretase complex, map the substrate initial docking site(s), and investigate if multiple docking sites exist on the gamma-secretase complex for different transmembrane substrates. This work should yield far more structural and mechanistic information on the unusual intramembrane enzyme, which is required for cell-fate decisions in all metazoans.

Public Health Relevance

Our proposed structural investigation of the gamma-secretase complex will yield better mechanistic understanding on the unusual intramembrane enzyme, which plays a key role in Alzheimer disease causing Abeta production, and is required for cell fate decision in all metazoans.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
5R01AG029979-02
Application #
8321960
Study Section
Biochemistry and Biophysics of Membranes Study Section (BBM)
Program Officer
Wise, Bradley C
Project Start
2011-09-01
Project End
2016-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
2
Fiscal Year
2012
Total Cost
$374,043
Indirect Cost
$169,043
Name
Brookhaven National Laboratory
Department
Type
DUNS #
027579460
City
Upton
State
NY
Country
United States
Zip Code
11973
Takeuchi, Hideyuki; Yu, Hongjun; Hao, Huilin et al. (2017) O-Glycosylation modulates the stability of epidermal growth factor-like repeats and thereby regulates Notch trafficking. J Biol Chem 292:15964-15973
Tian, Ye; Zhang, Yugang; Wang, Tong et al. (2016) Lattice engineering through nanoparticle-DNA frameworks. Nat Mater 15:654-61
Liu, Wenyan; Tagawa, Miho; Xin, Huolin L et al. (2016) Diamond family of nanoparticle superlattices. Science 351:582-6
Yu, Hongjun; Takeuchi, Hideyuki; Takeuchi, Megumi et al. (2016) Structural analysis of Notch-regulating Rumi reveals basis for pathogenic mutations. Nat Chem Biol 12:735-40
Yu, Hongjun; Takeuchi, Megumi; LeBarron, Jamie et al. (2015) Notch-modifying xylosyltransferase structures support an SNi-like retaining mechanism. Nat Chem Biol 11:847-54
Tian, Ye; Wang, Tong; Liu, Wenyan et al. (2015) Prescribed nanoparticle cluster architectures and low-dimensional arrays built using octahedral DNA origami frames. Nat Nanotechnol 10:637-44
Sun, Jingchuan; Shi, Yi; Georgescu, Roxana E et al. (2015) The architecture of a eukaryotic replisome. Nat Struct Mol Biol 22:976-82
Li, Huilin; Wolfe, Michael S; Selkoe, Dennis J (2009) Toward structural elucidation of the gamma-secretase complex. Structure 17:326-34
Osenkowski, Pamela; Li, Hua; Ye, Wenjuan et al. (2009) Cryoelectron microscopy structure of purified gamma-secretase at 12 A resolution. J Mol Biol 385:642-52