How to make membrane proteins and not unfold them

Originally Aired June 7, 2021
 

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How to make membrane proteins and not unfold them

Originally Aired June 7, 2021
 

Overview

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Membrane proteins represent more than 60 percent of pharmacological targets against human diseases, making them desirable targets for drug discovery campaigns. But, as this webcast will address, producing and purifying high-quality membrane proteins can be very challenging.

Membrane proteins are often intrinsically flexible and unstable, resulting in poor expression yield, solubility issues, and instability when recombinantly overexpressed. Introducing point mutations can reduce flexibility and increase stability without removing protein activity; however, finding stabilising mutations has traditionally been a painstaking trial and error process of testing at every amino acid position. Now, in silico and bioinformatic methods can help to predict positions likely to have a stabilising effect.

Membrane protein instability and flexibility issues are also often exacerbated when proteins are solubilised in detergent during purification. An emerging technology is the use of synthetic polymers for detergent-free extraction of protein. Rather than replacing native lipid with detergent, these polymers act as “cookie-cutters” to release membrane proteins in a belt of native lipids, resulting in increased activity and prolonged stability of the protein product.

You will learn:
  • How to recombinantly express, solubilize and purify membrane proteins
  • In silico methods to select stabilising point mutations, and how this can alleviate bottlenecks in membrane protein production
  • Detergent-free methods to extract membrane proteins and the benefits to purification and stability

This webcast has been produced by Peak Proteins, who retains sole responsibility for content. About this content.

Presenters

Presenter
Dr. Steven Harborne
Specialist in Membrane Protein Biochemistry
Peak Proteins, Cheshire UK
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Presenter
Dr. Alice Rothnie
Senior Lecturer in Biosciences
Aston University, Birmingham, UK
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Presenter
Moderator: Sarah Hiddleston
Science Journalist
Nature Research for Nature Middle East
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