Abstract
Mammalian DSPs have been historically isolated either from native tissue sources or from transfected insect cell cultures via time-consuming and cumbersome protocols often yielding protein of variable quality and quantity. A facile and highly reproducible alternative methodology involving the heterologous expression and purification of soluble mammalian DSPs from E. coli, which yields highly active and functional protein of a uniform quality and quantity, free of spurious posttranslational modifications inherent to mammalian and insect cell expression systems, is described in this chapter.
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Acknowledgement
This work was supported by National Institutes of Health grant R01GM121583 awarded to R. R.
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Stepanyants, N., Macdonald, P.J., Madan Mohan, P., Ramachandran, R. (2020). A Single Common Protocol for the Expression and Purification of Soluble Mammalian DSPs from Escherichia coli. In: Ramachandran, R. (eds) Dynamin Superfamily GTPases. Methods in Molecular Biology, vol 2159. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0676-6_3
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DOI: https://doi.org/10.1007/978-1-0716-0676-6_3
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