Expression and purification of active human receptor interacting protein 1 kinase using a baculovirus system
| dc.contributor.author | Maki, Jenny L. | |
| dc.contributor.author | Brazell, Tres | |
| dc.contributor.author | Cuny, Gregory D. | |
| dc.contributor.author | Degterev, Alexei | |
| dc.date.accessioned | 2020-03-10T17:31:03Z | |
| dc.date.available | 2020-03-10T17:31:03Z | |
| dc.date.issued | 2014-06 | |
| dc.description.abstract | Receptor Interacting Protein 1 (RIP1) kinase is one of the key mediators of tumor necrosis factor alpha (TNF-?) signaling and is critical for activation of necroptotic cell death. We developed a method for expression of recombinant kinase, utilizing baculovirus co-infection of Cdc37, an Hsp90 co-chaperone, and RIP1-His, followed by a two-step purification scheme. After optimization, 1–3 mg of highly purified RIP1 kinase was typically obtained from a 1 L of Sf9 cells. The recombinant protein displayed kinase activity that was blocked by RIP1 inhibitors, necrostatins. The purified protein was used to develop a simple and robust thermal shift assay for further assessment of RIP1 inhibitors. | |
| dc.identifier.citation | Copyright 2013 Protein Expression and Purification. This is a post-print version of a published paper that is available at: https://www.sciencedirect.com/science/article/pii/S1046592813000478. Recommended citation: Maki, Jenny L., J. Tres Brazell, Xin Teng, Gregory D. Cuny, and Alexei Degterev. "Expression and purification of active receptor interacting protein 1 kinase using a baculovirus system." Protein expression and purification 89, no. 2 (2013): 156-161.doi: 10.1016/j.pep.2013.03.002. This item has been deposited in accordance with publisher copyright and licensing terms and with the author's permission. | |
| dc.identifier.uri | https://hdl.handle.net/10657/5938 | |
| dc.language.iso | en_US | |
| dc.publisher | Protein Expression and Purification | |
| dc.subject | RIP1 kinase | |
| dc.subject | Cdc37 | |
| dc.subject | Necrostatins | |
| dc.subject | Baculovirus | |
| dc.subject | Thermal shift assay | |
| dc.title | Expression and purification of active human receptor interacting protein 1 kinase using a baculovirus system | |
| dc.type | Article |