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Mammalian SOD2 is exclusively located in mitochondria and not present in peroxisomes

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Abstract

Superoxide dismutases (SODs) are metalloenzymes that belong to the essential antioxidant enzyme systems of virtually all oxygen-respiring organisms. SODs catalyze the dismutation of highly reactive superoxide radicals into hydrogen peroxide and molecular oxygen. For the subcellular localization of the manganese superoxide dismutase (SOD2) in eukaryotic cells, a dual mitochondrial localization and peroxisomal localization were proposed in the literature. However, our own observation from immunofluorescence preparations of human and mouse tissues suggested that SOD2 serves as an excellent marker protein for mitochondria but never co-localized with peroxisomes. To clarify whether our observations were correct, we have carefully reinvestigated the subcellular localization of SOD2 using sensitive double-immunofluorescence methods on frozen and paraffin sections as well as in cell culture preparations. In addition, ultrastructural analyses were performed with post-embedding immunoelectron microscopy on LR White sections as well as labeling of ultrathin cryosections with various immunogold techniques. In all morphological experiments, the SOD2 localization was compared to one of the catalase, a typical marker protein for peroxisomes, solely localized in these organelles. Moreover, biochemical subcellular fractions of mouse liver was used to isolate enriched organelles and highly purified peroxisomal fractions for Western blot analyses of the exact subcellular distributions of SOD2 and catalase. All results with the various methodologies, tissues, and cell types used revealed that catalase and SOD2 were always confined to distinct and separate subcellular compartments. SOD2 was unequivocally in mitochondria, but never present in peroxisomes. Furthermore, our results are supported by accumulating database information on organelle proteomes that also indicate that SOD2 is a pure mitochondrial protein.

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Abbreviations

AECII:

Alveolar epithelial cell type II

PFA:

Paraformaldehyde

PBS:

Phosphate-buffered saline

SOD2:

Superoxide dismutase 2

CAT:

Catalase

ROS:

Reactive oxygen species

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Acknowledgments

The authors thank Dr. Klaus-Peter Valerius for help in animal perfusions. The excellent technical assistance of Elke Richter and Bianca Pfeiffer is gratefully acknowledged. We would like to thank Profs. Denis I. Crane for providing us the catalase antibody (see Table 1). Finally, we are indebted to Jessica Woods for carefully reading and correcting the manuscript.

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  1. Georg Lüers

    Present address: Institute for Anatomy and Experimental Morphology, UKE, Hamburg, Germany

Authors and Affiliations

  1. Division of Medical Cell Biology, Institute for Anatomy and Cell Biology II, Justus Liebig University, Aulweg 123, 35385, Giessen, Germany

    Srikanth Karnati, Georg Lüers, Susanna Pfreimer & Eveline Baumgart-Vogt

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  1. Srikanth Karnati
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  2. Georg Lüers
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Correspondence to Eveline Baumgart-Vogt.

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Karnati, S., Lüers, G., Pfreimer, S. et al. Mammalian SOD2 is exclusively located in mitochondria and not present in peroxisomes. Histochem Cell Biol 140, 105–117 (2013). https://doi.org/10.1007/s00418-013-1099-4

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