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Isolation of mitochondria with cubic membrane morphology reveals specific ionic requirements for the preservation of membrane structure

  • New Methods in Cell Biology
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Abstract

Biological membranes with cubic symmetry are a hallmark of virus-infected or diseased cells. The mechanisms of formation and specific cellular functions of cubic membranes, however, are unclear. The best-documented cubic membrane formation occurs in the free-living giant amoeba Chaos carolinense. In that system, mitochondrial inner membranes undergo a reversible structural change from tubular to cubic membrane organization upon starvation of the organism. As a prerequisite to further analyze the structural and functional features of cubic membranes, we adapted protocols for the isolation of mitochondria from starved amoeba and have identified buffer conditions that preserve cubic membrane morphology in vitro. The requirement for high concentration of ion-chelating agents in the isolation media supports the importance of a balanced ion milieu in establishing and maintaining cubic membranes in vivo.

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Acknowledgments

We would like to thank Ms. Shoon Meiyin, Ms. Low Chwee Wah, and Mr. Lim Teck Kwang for technical assistance. The electron microscopy and proteomics studies were conducted at the Electron Microscopy Unit at Yong Loo Lin School of Medicine, and the Proteins and Proteomics Center at the National University of Singapore, respectively. This work was supported by grants from the Austrian Science Funds, FWF (Project F3005 LIPOTOX) to S.D.K. and BMRC, Singapore (R-185-000-197-305) to Y.D.

Conflict of interest

The authors have no conflict of interest. All forms of financial support are acknowledged in the contribution. No agreement is signed with any sponsor of the research reported that forbids our publishing of this research without the prior approval of the sponsor.

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Authors and Affiliations

  1. Cubic Membrane Laboratory, Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 117597, Singapore

    Ketpin Chong, Olivia Li Ling Tan, Zakaria A. Almsherqi & Yuru Deng

  2. Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore, 117543, Singapore

    Qingsong Lin

  3. Institute of Molecular Biosciences, BioTechMed-Graz, University of Graz, Humboldtstrasse 50/II, 8010, Graz, Austria

    Sepp D. Kohlwein

  4. Institute of Biomedical Engineering and Health Sciences, Changzhou University, Changzhou, 213164, Jiangsu, China

    Yuru Deng

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  1. Ketpin Chong
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Correspondence to Yuru Deng.

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Chong, K., Tan, O.L.L., Almsherqi, Z.A. et al. Isolation of mitochondria with cubic membrane morphology reveals specific ionic requirements for the preservation of membrane structure. Protoplasma 252, 689–696 (2015). https://doi.org/10.1007/s00709-014-0698-9

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  • DOI: https://doi.org/10.1007/s00709-014-0698-9

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