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. Author manuscript; available in PMC: 2023 Dec 11.
Published in final edited form as: Curr Opin Struct Biol. 2020 Feb 15;60:150–156. doi: 10.1016/j.sbi.2020.01.006

Figure 2. Different strategy that have been successfully used for production of large nanodiscs.

Figure 2.

(a) Design of the Npu DnaE split-intein MSP fusion construct and mechanism of intein splicing in vivo, leading to the generation of circularized MSP. (b) A general outline of the constructs for making covalently circularized nanodiscs using sortase. Glycine (G) and LPXTG at the N and C termini respectively are necessary for the linkage by sortase. (c) DNA-corralled reconstitution strategy. Addition of detergent and lipids to small nanodisc- decorated barrels followed by dialysis results in the construction of a large bilayer nanodisc within the DNA cavity. Each DNA barrel is decorated with ssDNA overhang as handles to hybridize with ssDNA chemically conjugated small nanodisc.