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Review
. 2018 Aug:51:129-134.
doi: 10.1016/j.sbi.2018.03.014. Epub 2018 Apr 20.

Covalently circularized nanodiscs; challenges and applications

Mahmoud L Nasr  1 , Gerhard Wagner  2
Affiliations
Review

Covalently circularized nanodiscs; challenges and applications

Mahmoud L Nasr et al. Curr Opin Struct Biol. 2018 Aug.

Abstract

Covalently circularized nanodiscs (cNDs) represent a significant advance in the durability and applicability of nanodisc technology. The new cNDs demonstrate higher size homogeneity and improved stability compared with that of non-circularized forms. Moreover, cNDs can be prepared at various defined sizes up to 80-nm diameter. The large cNDs can house much larger membrane proteins and their complexes than was previously possible with the conventional nanodiscs. In order to experience the full advantages of covalent circularization, high quality circularized scaffold protein and nanodisc samples are needed. Here, we give a concise overview and discuss the technical challenges that needed to be overcome in order to obtain high quality preparations. Furthermore, we review some potential new applications for the cNDs.

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Conflict of interest statement

Conflict of Interest.

The authors have founded NOW Scientific Inc to allow purchase of assembled circularized MSPs. Expression plasmid are available for a nominal fee, and detailed protocols have been published.

Figures

Figure 1
Figure 1
Circularization and multimerization of NWs by sortase A. (a) The addition of sortase A to a concentrated scaffold protein solution can lead to multimerization followed by circularization. (b) A general outline of the constructs that are used for making covalently circularized nanodiscs. This figure is adapted from reference .
Figure 2
Figure 2
Schematic of the in solution method for scaffold protein circularization. After the IMAC step, cNWs are further purified by size-exclusion followed by ion exchange chromatography to remove the multimerization and hydrolysis byproducts.
See this image and copyright information in PMC

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