Tetrameric structure and cellular location of catechol 2,3-dioxygenase |
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| Authors: | Jörg Winkler Lindsay D Eltis Daryl F Dwyer Manfred Rohde |
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| Institution: | (1) Department of Microbiology, National Research Center for Biotechnology, D-38124 Braunschweig, Germany;(2) Department of Biochemistry, Université Laval, G1K 7P4 Ste-Foy, Québec, Canada;(3) Department of Civil and Mineral Engineering, University Minnesota, 55455 Minneapolis, MN, USA;(4) Present address: Gesellschaft für Biotechnologische Forschung mbH (GBF), Mascheroder Weg 1, D-38124 Braunschweig, Germany |
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| Abstract: | Catechol 2,3-dioxygenase from the meta-cleavage pathway encoded on the TOL plasmid of Pseudomonas putida (pWWO) was investigated by electron microscopy. Negatively stained samples of the purified catechol 2,3-dioxygenase revealed that the enzyme consists of four subunits arranged in a tetrahedral conformation. Monoclonal antibodies raised against catechol 2,3-dioxygenase showed highly specific reactions and were used to localize the enzyme in Escherichia coli (pAW31) and P. putida (pWWO), using the protein A-gold technique carried out as a post-embedding immunoelectron microscopy procedure. Our in situ labeling studies revealed a cytoplasmic location of the catechol 2,3-dioxygenase in both cell types.Abbreviations
C23O
Catechol 2,3-dioxygenase
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3MB 3
Methylbenzoate
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AK1
Anti-C23O-IgG-antibody
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G
Gold particle |
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| Keywords: | Catechol 2 3-dioxygenase Pseudomonas putida TOL plasmid Monoclonal antibody Immunoelectron microscopy Quaternary structure |
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