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Data publication: DNA-Mediated Stack Formation of Nanodiscs

Subramanian, Madhumalar; Kielar, Charlotte; Tsushima, Satoru; Fahmy, Karim; Oertel, Jana

The recent publication showed the DNA-mediated stack formation of nanodiscs. Background: Membrane-scaffolding proteins (MSPs) become a versatile tool in generating nano-sized discoidal membrane mimetics (nanodiscs) for membrane protein research. We describe here the formation of multimers of membrane-scaffolding protein MSP1D1-bounded nanodiscs using the thiol reactivity of engineered cysteines. The mutated positions N42 and K163 in MSP1D1 were chosen to support chemical modification as evidenced by fluorescent labeling with pyrene. The direct disulphide bond formation of nanodiscs formed by the MSP1D1_N42C variant led to dimers and trimers with low yield. In contrast, transmission electron microscopy revealed that the attachment of oligonucleotides to the engineered cysteines of MSP1D1 allowed the growth of submicron-sized tracts of stacked nanodiscs through the hybridization of nanodisc populations carrying complementary strands and a flexible spacer.

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