title: Tetrameric structure and cellular location of catechol 2,3-dioxygenase.
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.
authors.affiliation: Department of Microbiology, National Research Center for Biotechnology, Braunschweig, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Winkler
authors.forename: L D
authors.initials: LD
authors.lastname: Eltis
authors.forename: D F
authors.initials: DF
authors.lastname: Dwyer
authors.forename: M
authors.initials: M
authors.lastname: Rohde
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D050557:Catechol 2,3-Dioxygenase; D002396:Catechols / Q000378:metabolism; D003593:Cytoplasm / Q000201:enzymology; D049308:Dioxygenases; D004926:Escherichia coli / Q000201:enzymology* / Q000648:ultrastructure; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D010105:Oxygenases / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure; D011487:Protein Conformation; D016958:Pseudomonas putida / Q000201:enzymology* / Q000648:ultrastructure
chemical_list: D000911:Antibodies, Monoclonal; D002396:Catechols; D010105:Oxygenases; D049308:Dioxygenases; D050557:Catechol 2,3-Dioxygenase; C034221:catechol