Distribution of the flavohaemoglobin, HMP, between periplasm and cytoplasm in Escherichia coli |
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| Authors: | Subhash G Vasudevan Pan Tang Nicholas E Dixon Robert K Poole |
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| Institution: | Department of Molecular Sciences, James Cook University, Townsville, Qld 4811, Australia; Centre for Molecular Structure and Function, Research School of Chemistry, Australian National University, Canberra, Australia; Division of Life Sciences, King's College London, UK; Division of Biochemistry and Molecular Biology, John Curtin School of Medical Research, Australian National University, Canberra, Australia |
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| Abstract: | Abstract The subcellular distribution of the soluble flavohaemoglobin (HMP) of Escherichia coli has been determined. Cells over-expressing HMP from the cloned hmp gene on a multicopy plasmid were fractionated by osmotic shock and lysozyme treatment. Spectral analysis of subcellular fractions showed the CO-binding haemoprotein to be cytoplasmic. However, Western blotting using antibody raised to purified HMP revealed approximately 30% of the protein to be periplasmic in the over-expressing strain. Western analysis also revealed substantial levels of periplasmic HMP in a strain expressing only chromosomally encoded protein but none in an hmp mutant. The results are discussed in relation to protein function and the similar distribution reported for Vitreoscilla globin. |
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| Keywords: | Haemoglobin Globin-like proteins Cytoplasm Periplasm Oxygen utilisation Escherichia coli |
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