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Smaller Nanodiscs are Suitable for Studying Protein Lipid Interactions by Solution NMR

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Abstract

Phospholipid bilayer nanodiscs, a newly developed model membrane system, provides “native-like” membrane environment for membrane protein studies. Nanodiscs assembled by membrane scaffold protein and phospholipid bilayer, with defined sizes that can be accurately regulated by changing the amino acid residues of the MSP construct. Herein we described the expression and purification of ΔMSP, a deletion mutant of the membrane scaffold protein. Smaller nanodiscs with mixed lipids were assembled, and the observed 31P NMR spectra showed identical chemical shifts to those of nanodiscs with pure POPC and POPE lipids, indicating they share similar chemical environments. The success of incorporation STIM1-TM into nanodiscs indicated the application of this smaller nanodisc system can be used to membrane protein studies by solution NMR.

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Abbreviations

MSP:

Membrane scaffold protein

TEV:

Tobacco etch virus

Histag:

Hexahistidine affinity tag

IPTG:

Isopropyl-thio-galactoside

POPE:

1-Hexadecanoyl-2-(9Z-octadecenoyl)-sn-glycero-3-phosphoethanolamine

POPC:

1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine

DMPC:

1,2-Dimyristoyl-sn-glycero-3-phosphocholine

TPC:

N-tetradecylphosphocholine

DLS:

Dynamic light scattering

NMR:

Nuclear magnetic resonance

SDS–PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

HSQC:

Heteronuclear singular quantum correlation

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Acknowledgments

This work was supported by National Natural Science Foundation of China(No.31200579, National Key Basic Research Program from Ministry of Science and Technology (2012CB917202), and Hefei Center for Physical Science and Technology.

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Authors and Affiliations

  1. High Magnetic Field Laboratory, Hefei Institutes of Physical Science, Chinese Academy of Science, Hefei, 230031, Anhui, China

    Xiaoming Wang, Yan Li, Yunchen Bi & Yujuan Wang

  2. National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Chaoyang District, Beijing, 100101, China

    Zongjun Mu

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  1. Xiaoming Wang
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  2. Zongjun Mu
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  3. Yan Li
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  4. Yunchen Bi
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  5. Yujuan Wang
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Correspondence to Yujuan Wang.

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Wang, X., Mu, Z., Li, Y. et al. Smaller Nanodiscs are Suitable for Studying Protein Lipid Interactions by Solution NMR. Protein J 34, 205–211 (2015). https://doi.org/10.1007/s10930-015-9613-2

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