Cell-free expression and nanodiscs: New applications for stabilized membrane proteins

Nanodiscs are excellent tools for membrane protein stabilization. However, traditional methods require detergents for nanodisc complex assembly, which might affect membrane protein activity. Cell-free lysates combined with nanodiscs provide a detergent-free alternative to generate nanodisc-stabilized membrane proteins.

In the past few months, a number of exciting data have been published which extend the use of nanodiscs & cell-free lysates to new applications, and provide new insights.


  Nanodiscs and cell-free expression: the perfect match for functional membrane proteins

NMR: From nanodiscs to isotropic bicelles. Laguerre et al. addressed resolution issues in NMR commonly found with membrane proteins. Nanodiscs have been used as stabilizing tools for NMR analysis of membrane proteins. However, resolution was limited because of protein scaffold background signals. The authors developed a protocol to transfer membrane proteins from the nanodisc complex into bicelles, which greatly improved the quality of NMR spectra. Read more



Crystallization: From nanodiscs to meso phase. Crystallization was believed to be a difficult application for membrane proteins stabilized in nanodiscs. Nikolaev et al. were able to show that bacteriorhodopsin:nanodisc complexes can yield BR crystals in a meso phase setup. Read more



LILBID MS: New insights to protein oligomerization Henrich et al. used non-covalent mass spectrometry to answer the most frequently asked questions about nanodiscs:

- How many membrane scaffold proteins and how many lipids are in a nanodisc?

- What is the size of a nanodisc?

- Can membrane proteins be present as oligomers in nanodiscs? And how can this be detected?

In addition, they were able to show the effect of lipid composition on dimerization and activity of bacterial MraY, an important antimicrobial target. Read more



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Literature references:
Laguerre, A. et al. From nanodiscs to isotropic bicelles: A procedure for solution nuclear magnetic resonance studies of detergent-sensitive integral membrane proteins. Structure (2016) 24, 1-12.
Nikolaev, M. et al. Integral membrane proteins can be crystallized directly from nanodiscs. Cryst. Growth Des. (2017) 17(3), 945–948 
Henrich, E., et al. Analyzing native membrane protein assembly in nanodiscs by combined non-covalent mass spectrometry and synthetic biology. eLife (2017) 6:e20954.