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Issue 16, 2019, Issue in Progress
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Doubly spin-labeled nanodiscs to improve structural determination of membrane proteins by ESR

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Abstract

Pulsed dipolar spectroscopy (PDS) is a powerful tool to explore conformational changes of membrane proteins (MPs). However, the MPs suffer from relatively weak dipolar signals due to their complex nature in membrane environments, which consequently reduces the interspin distance resolution obtainable by PDS. Here we report the use of nanodiscs (NDs) to improve the distance resolution. Two genetically engineered membrane scaffold protein mutants are introduced, each of which is shown to form double-labeled ND efficiently and with high homogeneity. The resultant interspin distance distribution is featured by a small distribution width, suggesting high resolution. When PDS is performed on a binary mixture of the double-labeled ND devoid of MPs and the un-labeled ND with incorporated double-labeled MPs, the overall amplitude of dipolar signals is increased, leading to a critical enhancement of the distance resolution. A theoretical foundation is provided to validate the analysis. With this approach, the determination of MP structures can be studied at high resolution in NDs.

Graphical abstract: Doubly spin-labeled nanodiscs to improve structural determination of membrane proteins by ESR

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Publication details

The article was received on 02 Feb 2019, accepted on 14 Mar 2019 and first published on 19 Mar 2019


Article type: Paper
DOI: 10.1039/C9RA00896A
RSC Adv., 2019,9, 9014-9021
  • Open access: Creative Commons BY-NC license
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    Doubly spin-labeled nanodiscs to improve structural determination of membrane proteins by ESR

    C. Li, C. Hung, P. Yeh, C. Li and Y. Chiang, RSC Adv., 2019, 9, 9014
    DOI: 10.1039/C9RA00896A

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