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Structural characterization of the human membrane protein VDAC2 in lipid bilayers by MAS NMR

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Abstract

The second isoform of the human voltage dependent anion channel (VDAC2) is a mitochondrial porin that translocates calcium and other metabolites across the outer mitochondrial membrane. VDAC2 has been implicated in cardioprotection and plays a critical role in a unique apoptotic pathway in tumor cells. Despite its medical importance, there have been few biophysical studies of VDAC2 in large part due to the difficulty of obtaining homogeneous preparations of the protein for spectroscopic characterization. Here we present high resolution magic angle spinning nuclear magnetic resonance (NMR) data obtained from homogeneous preparation of human VDAC2 in 2D crystalline lipid bilayers. The excellent resolution in the spectra permit several sequence-specific assignments of the signals for a large portion of the VDAC2 N-terminus and several other residues in two- and three-dimensional heteronuclear correlation experiments. The first 12 residues appear to be dynamic, are not visible in cross polarization experiments, and they are not sufficiently mobile on very fast timescales to be visible in 13C INEPT experiments. A comparison of the NMR spectra of VDAC2 and VDAC1 obtained from highly similar preparations demonstrates that the spectral quality, line shapes and peak dispersion exhibited by the two proteins are nearly identical. This suggests an overall similar dynamic behavior and conformational homogeneity, which is in contrast to two earlier reports that suggested an inherent conformational heterogeneity of VDAC2 in membranes. The current data suggest that the sample preparation and spectroscopic methods are likely applicable to studying other human membrane porins, including human VDAC3, which has not yet been structurally characterized.

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Acknowledgements

This research was supported by NIH Grants EB001960 and EB002026. MTE acknowledges support from an American Cancer Society postdoctoral research fellowship. The authors thank Mr. Frans Ricardo for MALDI mass spectrometry characterization of VDAC2.

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Author notes

  1. Matthew T. Eddy

    Present address: Departments of Chemistry, University of Florida, Gainesville, FL, 32611, USA

  2. Tsyr-Yan Yu

    Present address: Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, 10617, Taiwan, Republic of China

  3. Matthew T Eddy and Tsyr-Yan Yu have contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA

    Matthew T. Eddy & Robert G. Griffin

  2. Francis Bitter Magnet Laboratory, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA

    Matthew T. Eddy & Robert G. Griffin

  3. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, 02115, USA

    Tsyr-Yan Yu & Gerhard Wagner

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  1. Matthew T. Eddy
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Correspondence to Robert G. Griffin.

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Eddy, M.T., Yu, TY., Wagner, G. et al. Structural characterization of the human membrane protein VDAC2 in lipid bilayers by MAS NMR. J Biomol NMR 73, 451–460 (2019). https://doi.org/10.1007/s10858-019-00242-8

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  • DOI: https://doi.org/10.1007/s10858-019-00242-8

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