Efficient production of a mature and functional gamma secretase protease

I. Khan; S. Krishnaswamy; M. Sabale; D. Groth; L. Wijaya; M. Morici; I. Berger; C. Schaffitzel; P.E. Fraser; R.N. Martins; G. Verdile

doi:10.1038/s41598-018-30788-w

Back

Efficient production of a mature and functional gamma secretase protease

Journal article

Open access

Peer reviewed

Efficient production of a mature and functional gamma secretase protease

I. Khan, S. Krishnaswamy, M. Sabale, D. Groth, L. Wijaya, M. Morici, I. Berger, C. Schaffitzel, P.E. Fraser, R.N. Martins, …

Scientific Reports, Vol.8(1), art. no. 12834

2018

DOI: https://doi.org/10.1038/s41598-018-30788-w

Files and links (2)

pdf

Visualizing_Degradation_of_Black_Phosphorus.pdfDownload View

Published (Version of Record) Open Access

url

Free to Read *No subscription requiredView

Abstract

Baculoviral protein expression in insect cells has been previously used to generate large quantities of a protein of interest for subsequent use in biochemical and structural analyses. The MultiBac baculovirus protein expression system has enabled, the use of a single baculovirus to reconstitute a protein complex of interest, resulting in a larger protein yield. Using this system, we aimed to reconstruct the gamma (γ)-secretase complex, a multiprotein enzyme complex essential for the production of amyloid-β (Aβ) protein. A MultiBac vector containing all components of the γ-secretase complex was generated and expression was observed for all components. The complex was active in processing APP and Notch derived γ-secretase substrates and proteolysis could be inhibited with γ-secretase inhibitors, confirming specificity of the recombinant γ-secretase enzyme. Finally, affinity purification was used to purify an active recombinant γ-secretase complex. In this study we demonstrated that the MultiBac protein expression system can be used to generate an active γ-secretase complex and provides a new tool to study γ-secretase enzyme and its variants.

Details

Title: Efficient production of a mature and functional gamma secretase protease
Authors/Creators: I. Khan (Author/Creator) - Curtin University
S. Krishnaswamy (Author/Creator) - Edith Cowan University
M. Sabale (Author/Creator) - Curtin University
D. Groth (Author/Creator) - Curtin University
L. Wijaya (Author/Creator) - Edith Cowan University
M. Morici (Author/Creator) - The University of Western Australia
I. Berger (Author/Creator) - University of Bristol
C. Schaffitzel (Author/Creator) - University of Bristol
P.E. Fraser (Author/Creator) - University of Toronto
R.N. Martins (Author/Creator) - Edith Cowan University
G. Verdile (Author/Creator) - Curtin University
Publication Details: Scientific Reports, Vol.8(1), art. no. 12834
Publisher: Nature Publishing Group
Identifiers: 991005540882507891
Copyright: © 2018, The Author(s)
Murdoch Affiliation: School of Psychology and Exercise Science
Language: English
Resource Type: Journal article

UN Sustainable Development Goals (SDGs)

This output has contributed to the advancement of the following goals:

Source: InCites

Metrics

162 File views/ downloads

61 Record Views

4 Times Cited - Web of Science

InCites Highlights

These are selected metrics from InCites Benchmarking & Analytics tool, related to this output

Collaboration types: Domestic collaboration; International collaboration
Citation topics: 1 Clinical & Life Sciences; 1.52 Neurodegenerative Diseases; 1.52.57 Alzheimer's Mechanisms
Web Of Science research areas: Biochemistry & Molecular Biology
ESI research areas: Biology & Biochemistry