期刊
JOURNAL OF CELL SCIENCE
卷 128, 期 3, 页码 589-598出版社
COMPANY OF BIOLOGISTS LTD
DOI: 10.1242/jcs.164384
关键词
gamma-secretase; Alzheimer disease; Conformational landscape; Regulated intramembrane proteolysis; Single particle electron microscopy
类别
资金
- Fund for Scientific Research Flanders (FWO)
- research fund KU Leuven
- Hercules Foundation
- Federal Office for Scientific Affairs [IAP P7/16]
- Flemish Government
- Flanders Institute for Biotechnology (VIB)
- Agency for Innovation by Science and Technology (IWT)
- European Research Council (AdG)
- Marie Curie fellowship
- European Molecular Biology Organization (EMBO)
- Foundation for Alzheimer Research (SAO-FRA)
- National Institutes of Health [AG15379]
The structure and function of the gamma-secretase proteases are of great interest because of their crucial roles in cellular and disease processes. We established a novel purification protocol for the gamma-secretase complex that involves a conformation-and complexspecific nanobody, yielding highly pure and active enzyme. Using single particle electron microscopy, we analyzed the gamma-secretase structure and its conformational variability. Under steady-state conditions, the complex adopts three major conformations, which differ in overall compactness and relative position of the nicastrin ectodomain. Occupancy of the active or substrate-binding sites by inhibitors differentially stabilizes subpopulations of particles with compact conformations, whereas a mutation linked to familial Alzheimer disease results in enrichment of extended-conformation complexes with increased flexibility. Our study presents the gamma-secretase complex as a dynamic population of interconverting conformations, involving rearrangements at the nanometer scale and a high level of structural interdependence between subunits. The fact that protease inhibition or clinical mutations, which affect amyloid beta (A beta) generation, enrich for particular subpopulations of conformers indicates the functional relevance of the observed dynamic changes, which are likely to be instrumental for highly allosteric behavior of the enzyme.
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