Coproporphyrinogen III oxidase from barley and tobacco--sequence analysis and initial expression studies.
Coproporphyrinogen III oxidase (coprogen oxidase; EC 1.3.3.3) is part of the pathway from 5-amino-levulinate to protoporphyrin IX which is common in all organisms and catalyses oxidative decarboxylation at two tetrapyrrole side chains. We cloned and sequenced full-length cDNAs encoding coprogen oxidase from barley (Hordeum vulgare L.) and tobacco (Nicotiana tabacum L.). They code for precursor peptides of 43.6 kDa and 44.9 kDa, respectively. Import into pea plastids resulted in a processed tobacco protein of approx. 39 kDa, which accumulated in the stroma fraction. Induction of synthesis of recombinant putative tobacco mature coprogen oxidase consisting of 338 amino-acid residues in Escherichia coli at 20 degrees C result in a catalytically active protein of approx. 39 kDa, while induction of its formation at 37 degrees C immediately terminated bacterial growth, possibly due to toxic effects on the metabolic balance of tetrapyrrole biosynthesis. The plant coprogen oxidase gene was expressed to different extents in all tissues investigated. This is most likely due to the differing requirements for tetrapyrroles in different organs. The steady-state level of mRNA did not significantly differ in etiolated and greening barley leaves. The content of coprogen oxidase RNA reached its maximum in developing cells and decreased drastically when cells were completely differentiated. Functioning of the two photosystems apparatus requires the synthesis of all pigment and protein components during plant development. It is speculated that the enzymes involved in tetrapyrrole synthesis are developmentally rather than light-dependently regulated. Regulation of these enzymes also guarantees a constant flux of metabolic intermediates and avoids photodynamic damage by accumulating porphyrins.
Kruse
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Institut für Pflanzengenetik und Kulturpflanzenforschung, Gatersleben, Germany.
Mock
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HP
Grimm
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B
D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D002736:Chloroplasts / Q000201:enzymology; D003001:Cloning, Molecular; D003304:Coproporphyrinogen Oxidase / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary; D004792:Enzyme Precursors / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D001467:Hordeum / Q000201:enzymology*; D008027:Light; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D012333:RNA, Messenger / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D014026:Nicotiana / Q000201:enzymology*
D018076:DNA, Complementary; D004792:Enzyme Precursors; D012333:RNA, Messenger; D003304:Coproporphyrinogen Oxidase