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Coproporphyrinogen III oxidase from barley and tobacco — sequence analysis and initial expression studies

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Abstract

Coproporphyrinogen III oxidase (coprogen oxidase; EC 1.3.3.3) is part of the pathway from 5-aminolevulinate to protoporphyrin IX which is common in all organisms and catalyses oxidative decarboxylation at two tetrapyrrole side chains. We cloned and sequenced fulllength 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°C result in a catalytically active protein of approx. 39 kDa, while induction of its formation at 37°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.

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Abbreviations

ALA:

5-aminolevulinic acid

copro:

coproporphyrin III

coprogen:

coproporphyrinogen III

IPTG:

isopropyl β-d-thiogalactopyranoside

LHCP II:

light-harvesting chlorophyll-binding protein of photosystem II

OD:

optical density

proto IX:

protoporphyrin IX

protogen IX:

protoporphyrinogen IX

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Authors and Affiliations

  1. Institut für Pflanzengenetik und Kulturpflanzenforschung, Corrensstrasse 3, D-06466, Gatersleben, Germany

    Elisabeth Kruse, Hans-Peter Mock & Bernhard Grimm

Authors
  1. Elisabeth Kruse
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  2. Hans-Peter Mock
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  3. Bernhard Grimm
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Additional information

Accession number: The nucleotide sequence data reported will appear in the EMBL, GenBank and DDBJ Nucleotide Sequence Databases under the accession numbers X82830 (barley coprogen oxidase) and X82831 (tobacco coprogen oxidase).

We are grateful to Prof. Marvin Smith (IPK, Gatersleben, Germany) for his valuable suggestions on earlier versions of the manuscript and to the Ministerium für Wissenschaft und Forschung des Landes Sachsen-Anhalt for financial support to B. Grimm. We thank Petra Linow for skillful technical assistance, Dr. Sakari Kauppinen (Carlsberg Laboratory, Copenhagen, Denmark) for providing the barley cDNA library and Dr. Ole Madsen for providing the soybean coprogen oxidase cDNA clone.

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Kruse, E., Mock, HP. & Grimm, B. Coproporphyrinogen III oxidase from barley and tobacco — sequence analysis and initial expression studies. Planta 196, 796–803 (1995). https://doi.org/10.1007/BF00197347

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  • DOI: https://doi.org/10.1007/BF00197347

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