Generation and purification of submitochondrial particles of different polarities from plant mitochondria

FEBS Letters

Ericson, Ingemar³  Gardestrom, Per²  Møller, Ian M.⁴  Kay, Christopher J.¹  Palmer, John M.¹ 

[1] Department of Pure and Applied Biology, Imperial College of Science and Technology, Prince Consort Road, London SW7 2BB, England;Department of Plant Physiology, University of Umeå, S-901 87 UmeåSweden;Department of Biochemistry, University of Umeå, S-901 87 UmeåSweden;Department of Plant Physiology, University of Lund, Box 7007, S-220 07 Lund, Sweden

关键词: Mitochondria;    (Plant);    Polarity;    Sidedness;    Submitochondrial particle;    Two-phase partitioning;    HMDA;    hexamethylenediamine;    IO;    inside-out;    MDH;    malate dehydrogenase;    Mops;    4-morpholinepropanesulfonic acid;    RO;    right-side-out;    SDK;    succinate dehydrogenase;    SMP;    submitochondrial particle(s);   

DOI  :  10.1016/0014-5793(85)80144-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Mitochondria were isolated from Jerusalem artichoke (Helianthus tuberosus L.) tubers in a low-salt medium. Submitochondrial particles (SMP) produced by sonication in a low salt medium + 20 mM MgCl₂ were 18% right-side-out (RO) as judged by the latency of cytochrome c oxidase assayed ± Triton X-100. SMP produced by French press treatment in a low-salt medium + 5 mM EDTA to remove bound divalent cations were 98% RO. Less extreme treatments gave SMP of intermediate polarity. There was a positive correlation between the % RO-SMP (produced by sonication) and the % NAD⁺-malate dehydrogenase enclosed within the SMP indicating that only RO-SMP contained trapped matrix. When a mixed population of SMP (45% RO) was applied to an aqueous polymer two-phase system, the top phase contained 76% RO-SMP and the bottom phase mostly inside-out SMP (26% RO). By analogy with the models for stacking of photosynthetic membranes, we propose that crista formation in the inner mitochondrial membrane is electrostatically regulated and SMP deriving from the closely stacked crista regions are inside-out.

【 授权许可】

Unknown