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Method Article
Naotada Ishihara
Ishihara lab, Kurume University
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
This protocol describes a methods for expression and purification of human recombinant L-OPA1 using the BmNPV bacmid-silkworm expression system. Using this method, we have shown how the dynamin-like GTPase and membrane lipids mediate membrane fusion in mammals in associated publication ”Molecular basis of selective mitochondrial fusion by heterotypic action between OPA1 and cardiolipin” in Nat. Cell Biol..
Because bacmid preparation and silkworm culture is easy, and the bacmid-silkworm expression system might have advantages for the expression of large membrane proteins in comparison with Escherichia coli expression system. the present method will have general applicability to various mitochondrial membrane proteins. It takes 5 days for bacmid preparation, 7 days for protein expression, 2 days for protein purification and 1 days for detergent exchange, respectively.
Keywords
mitochondria, membrane fusion, dynamin-like GTPase, BmNPV bacmid-silkworm expression system
The authors declare no competing financial interests.
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Associated Publications
Molecular basis of selective mitochondrial fusion by heterotypic action between OPA1 and cardiolipin
by Tadato Ban, Takaya Ishihara, Hiroto Kohno, +6
Nature Cell Biology (20 June, 2017)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Naotada Ishihara
Ishihara lab, Kurume University
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 20 Jun, 2017
Community comments: 1
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
This protocol describes a methods for expression and purification of human recombinant L-OPA1 using the BmNPV bacmid-silkworm expression system. Using this method, we have shown how the dynamin-like GTPase and membrane lipids mediate membrane fusion in mammals in associated publication ”Molecular basis of selective mitochondrial fusion by heterotypic action between OPA1 and cardiolipin” in Nat. Cell Biol..
Because bacmid preparation and silkworm culture is easy, and the bacmid-silkworm expression system might have advantages for the expression of large membrane proteins in comparison with Escherichia coli expression system. the present method will have general applicability to various mitochondrial membrane proteins. It takes 5 days for bacmid preparation, 7 days for protein expression, 2 days for protein purification and 1 days for detergent exchange, respectively.
The authors declare no competing financial interests.
Loading...
Associated Publications
Molecular basis of selective mitochondrial fusion by heterotypic action between OPA1 and cardiolipin
by Tadato Ban, Takaya Ishihara, Hiroto Kohno, +6
Nature Cell Biology (20 June, 2017)