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Unusual location and characterization of Cu/Zn-containing superoxide dismutase from filamentous fungus Humicola lutea

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Abstract

The present study aims to provide new information about the unusual location of Cu/Zn-superoxide dismutase (Cu/Zn-SOD) in lower eukaryotes such as filamentous fungi. Humicola lutea, a high producer of SOD was used as a model system. Subcellular fractions [cytosol, mitochondrial matrix, and intermembrane space (IMS)] were isolated and tested for purity using activity measurements of typical marker enzymes. Evidence, based on electrophoretic mobility, sensitivity to KCN and H2O2 and immunoblot analysis supports the existence of Cu/Zn-SOD in mitochondrial IMS, and the Mn-SOD in the matrix. Enzyme activity is almost equally partitioned between both the compartments, thus suggesting that the intermembrane space could be one of the major sites of exposure to superoxide anion radicals. The mitochondrial Cu/Zn-SOD was purified and compared with the previously published cytosolic enzyme. They have identical molecular mass, cyanide- and H2O2-sensitivity, N-terminal amino acid sequence, glycosylation sites and carbohydrate composition. The H. lutea mitochondrial Cu/Zn-SOD is the first identified naturally glycosylated enzyme, isolated from IMS. These findings suggest that the same Cu/Zn-SOD exists in both the mitochondrial IMS and cytosol.

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Acknowledgments

This work was supported by the NCSI of the Ministry of Education and Science, Bulgaria (grant K-1302/02), which is greatly acknowledged.

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Authors and Affiliations

  1. The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, 26 Academician G. Bonchev, 1113, Sofia, Bulgaria

    Ekaterina Krumova, Svetlana Pashova, Lilyana Stefanova & Maria Angelova

  2. Institute of Organic Chemistry, Bulgarian Academy of Sciences, 9 Academician G. Bonchev, 1113, Sofia, Bulgaria

    Alexander Dolashki, Pavlina Dolashka-Angelova & Rumyana Hristova

  3. Interfacultary Institute of Biochemistry of Tuebingen University, Hoppe-Seyler-Straße 4, 72076, Tübingen, Germany

    Alexander Dolashki & Wolfgang Voelter

  4. Department of Immunology, Institute for Cell Biology, University of Tübingen, Auf der Morgenstelle 15, 72076, Tübingen, Germany

    Stefan Stevanovic

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  1. Ekaterina Krumova
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  2. Alexander Dolashki
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  3. Svetlana Pashova
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  4. Pavlina Dolashka-Angelova
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  9. Maria Angelova
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Corresponding author

Correspondence to Maria Angelova.

Additional information

Communicated by Geoffrey Turner.

Ekaterina Krumova and Alexander Dolashki equally contributed to this work.

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Krumova, E., Dolashki, A., Pashova, S. et al. Unusual location and characterization of Cu/Zn-containing superoxide dismutase from filamentous fungus Humicola lutea . Arch Microbiol 189, 121–130 (2008). https://doi.org/10.1007/s00203-007-0300-3

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  • DOI: https://doi.org/10.1007/s00203-007-0300-3

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