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Production of active glycosylation-deficient γ-secretase complex for crystallization studies

Andrés Ricardo López, Mitko Dimitrov, Hermeto Gerber, Virginie Braman, David L Hacker, Florian M Wurm, Patrick C Fraering
Alzheimer's disease (AD)-associated γ-secretase is a ubiquitously expressed multi-subunit protease complex embedded in the lipid bilayer of cellular compartments including endosomes and the plasma membrane. Although γ-secretase is of crucial interest for AD drug discovery, its atomic structure remains unresolved. γ-Secretase assembly and maturation is a multistep process, which includes extensive glycosylation on nicastrin (NCT), the only γ-secretase subunit having a large extracellular domain. These posttranslational modifications lead to protein heterogeneity that likely prevents the three-dimensional (3D) crystallization of the protease complex...
December 2015: Biotechnology and Bioengineering
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