Efficient production of a mature and functional gamma secretase protease
dc.contributor.author | Khan, I. | |
dc.contributor.author | Krishnaswamy, S. | |
dc.contributor.author | Sabale, M. | |
dc.contributor.author | Groth, David | |
dc.contributor.author | Wijaya, L. | |
dc.contributor.author | Morici, M. | |
dc.contributor.author | Berger, I. | |
dc.contributor.author | Schaffitzel, C. | |
dc.contributor.author | Fraser, P. | |
dc.contributor.author | Martins, R. | |
dc.contributor.author | Verdile, Giuseppe | |
dc.date.accessioned | 2018-12-13T09:12:20Z | |
dc.date.available | 2018-12-13T09:12:20Z | |
dc.date.created | 2018-12-12T02:46:29Z | |
dc.date.issued | 2018 | |
dc.identifier.citation | Khan, I. and Krishnaswamy, S. and Sabale, M. and Groth, D. and Wijaya, L. and Morici, M. and Berger, I. et al. 2018. Efficient production of a mature and functional gamma secretase protease. Scientific Reports. 8 (1): Article ID 12834. | |
dc.identifier.uri | http://hdl.handle.net/20.500.11937/72085 | |
dc.identifier.doi | 10.1038/s41598-018-30788-w | |
dc.description.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. | |
dc.publisher | Nature Publishing Group | |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | |
dc.title | Efficient production of a mature and functional gamma secretase protease | |
dc.type | Journal Article | |
dcterms.source.volume | 8 | |
dcterms.source.number | 1 | |
dcterms.source.issn | 2045-2322 | |
dcterms.source.title | Scientific Reports | |
curtin.department | School of Pharmacy and Biomedical Sciences | |
curtin.accessStatus | Open access |