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Lon protease preferentially degrades oxidized mitochondrial aconitase by an ATP-stimulated mechanism

Nature Cell Biology volume 4, pages 674–680 (2002)Cite this article

Abstract

Mitochondrial aconitase is sensitive to oxidative inactivation and can aggregate and accumulate in many age-related disorders. Here we report that Lon protease, an ATP-stimulated mitochondrial matrix protein, selectively recognizes and degrades the oxidized, hydrophobic form of aconitase after mild oxidative modification, but that severe oxidation results in aconitase aggregation, which makes it a poor substrate for Lon. Similarly, a morpholino oligodeoxynucleotide directed against the lon gene markedly decreases the amount of Lon protein, Lon activity and aconitase degradation in WI-38 VA-13 human lung fibroblasts and causes accumulation of oxidatively modified aconitase. The ATP-stimulated Lon protease may be an essential defence against the stress of life in an oxygen environment. By recognizing minor oxidative changes to protein structure and rapidly degrading the mildly modified protein, Lon protease may prevent extensive oxidation, aggregation and accumulation of aconitase, which could otherwise compromise mitochondrial function and cellular viability. Aconitase is probably only one of many mitochondrial matrix proteins that are preferentially degraded by Lon protease after oxidative modification.

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Figure 1: Loss of aconitase activity and protein modification after H2O2 treatment.
Figure 2: Degradation of oxidatively-denatured aconitase during Incubation with bovine heart muscle mitochondrial extract.
Figure 3: Elution profile of Lon protease from size-exclusion chromatography and proteolytic activity of bovine heart mitochondrial matrix fractions.
Figure 4: Purification of mitochondrial Lon protease.
Figure 5: Purified Lon protease preferentially degrades oxidized aconitase.
Figure 6: Diminished ATP-stimulated degradation of aconitase in intact WI-38 VA-13 human fibroblasts after treatment with lon antisense oligonucleotides.
Figure 7: Aconitase protein quantity, activity and oxidative modification in WI-38 VA-13 human lung fibroblasts treated with lon antisense oligonucleotides.

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Acknowledgements

This work was partially supported by NIH/NIEHS grant no. ES03598 and by NIH/NIA grant no. AG16256 to K.J.A.D.

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  1. Division of Molecular and Computational Biology, Ethel Percy Andrus Gerontology Center, University of Southern California, Los Angeles, 90089-0191, California, USA

    Daniela A. Bota & Kelvin J. A. Davies

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Correspondence to Kelvin J. A. Davies.

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Bota, D., Davies, K. Lon protease preferentially degrades oxidized mitochondrial aconitase by an ATP-stimulated mechanism. Nat Cell Biol 4, 674–680 (2002). https://doi.org/10.1038/ncb836

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