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A Single Common Protocol for the Expression and Purification of Soluble Mammalian DSPs from Escherichia coli

  • Natalia Stepanyants
  • Patrick J. Macdonald
  • Pooja Madan Mohan
  • Rajesh RamachandranEmail author
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Part of the Methods in Molecular Biology book series (MIMB, volume 2159)

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.

Key words

DSP Purification Escherichia coli Polyhistidine Tag Dynamin Drp1 OPA1 

Notes

Acknowledgement

This work was supported by National Institutes of Health grant R01GM121583 awarded to R. R.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Natalia Stepanyants
    • 1
  • Patrick J. Macdonald
    • 1
  • Pooja Madan Mohan
    • 2
  • Rajesh Ramachandran
    • 1
    • 3
    Email author
  1. 1.Department of Physiology and BiophysicsCase Western Reserve University School of MedicineClevelandUSA
  2. 2.Department of BiochemistryCase Western Reserve University School of MedicineClevelandUSA
  3. 3.Cleveland Center for Membrane and Structural BiologyCase Western Reserve University School of MedicineClevelandUSA

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