The syringaldazine-oxidizing peroxidase PXP 3-4 from poplar xylem: cDNA isolation, characterization and expression
- Author
- Jørgen Holst Christensen (UGent) , Serge Overney, Antje Rohde (UGent) , Wilson Ardiles Diaz (UGent) , Guy Bauw (UGent) , Patrice Simon, Marc Van Montagu (UGent) and Wout Boerjan (UGent)
- Organization
- Abstract
- The cell wall polymer lignin is believed to be condensed by specific cell wall-localized oxidoreductases. In many plants species, including poplar, the peroxidase-directed oxidation of the lignin analogue syringaldazine (SYR) has been localized to cells that undergo secondary wall formation, a process that includes lignification. As a first step to analyse the corresponding peroxidases, we have isolated previously two anionic isoenzymes (PXP 3-4 and PXP 5) from poplar xylem (Populus trichocarpa), which use SYR as a substrate. Here, we demonstrate that these enzymes are responsible for the visualized SYR oxidation in the developing xylem. The cDNA that corresponds to PXP 3-4 was isolated and the deduced protein was found closely related to the other SYR-oxidizing peroxidase PXP 5 (ca. 98% of identity). PXP 3-4 was expressed in a baculovirus expression system yielding high levels of active peroxidase (3 mg/l medium). The heterologously produced protein showed characteristics similar to those of the corresponding protein from poplar xylem (enzymatic properties, isoelectric point, and migration in a native gel). PXP 3-4 was expressed in the stem and in the root xylem. The data demonstrate that PXP 3-4 (and/or PXP 5) are present in differentiating xylem, supporting a function in secondary cell wall formation.
- Keywords
- MOLECULAR-CLONING, SUSPENSION-CULTURES, ANIONIC PEROXIDASE, POPULUS-KITAKAMIENSIS, HORSERADISH-PEROXIDASE, syringaldazine oxidation, secondary cell wall formation, poplar xylem peroxidase, lignification, cDNA isolation, expression analysis, AMINO-ACID, SEQUENCE, LIGNIFICATION, TOBACCO, LIGNIN
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-138599
- MLA
- Christensen, Jørgen Holst, et al. “The Syringaldazine-Oxidizing Peroxidase PXP 3-4 from Poplar Xylem: CDNA Isolation, Characterization and Expression.” PLANT MOLECULAR BIOLOGY, vol. 47, no. 5, 2001, pp. 581–93, doi:10.1023/A:1012271729285.
- APA
- Christensen, J. H., Overney, S., Rohde, A., Ardiles Diaz, W., Bauw, G., Simon, P., … Boerjan, W. (2001). The syringaldazine-oxidizing peroxidase PXP 3-4 from poplar xylem: cDNA isolation, characterization and expression. PLANT MOLECULAR BIOLOGY, 47(5), 581–593. https://doi.org/10.1023/A:1012271729285
- Chicago author-date
- Christensen, Jørgen Holst, Serge Overney, Antje Rohde, Wilson Ardiles Diaz, Guy Bauw, Patrice Simon, Marc Van Montagu, and Wout Boerjan. 2001. “The Syringaldazine-Oxidizing Peroxidase PXP 3-4 from Poplar Xylem: CDNA Isolation, Characterization and Expression.” PLANT MOLECULAR BIOLOGY 47 (5): 581–93. https://doi.org/10.1023/A:1012271729285.
- Chicago author-date (all authors)
- Christensen, Jørgen Holst, Serge Overney, Antje Rohde, Wilson Ardiles Diaz, Guy Bauw, Patrice Simon, Marc Van Montagu, and Wout Boerjan. 2001. “The Syringaldazine-Oxidizing Peroxidase PXP 3-4 from Poplar Xylem: CDNA Isolation, Characterization and Expression.” PLANT MOLECULAR BIOLOGY 47 (5): 581–593. doi:10.1023/A:1012271729285.
- Vancouver
- 1.Christensen JH, Overney S, Rohde A, Ardiles Diaz W, Bauw G, Simon P, et al. The syringaldazine-oxidizing peroxidase PXP 3-4 from poplar xylem: cDNA isolation, characterization and expression. PLANT MOLECULAR BIOLOGY. 2001;47(5):581–93.
- IEEE
- [1]J. H. Christensen et al., “The syringaldazine-oxidizing peroxidase PXP 3-4 from poplar xylem: cDNA isolation, characterization and expression,” PLANT MOLECULAR BIOLOGY, vol. 47, no. 5, pp. 581–593, 2001.
@article{138599, abstract = {{The cell wall polymer lignin is believed to be condensed by specific cell wall-localized oxidoreductases. In many plants species, including poplar, the peroxidase-directed oxidation of the lignin analogue syringaldazine (SYR) has been localized to cells that undergo secondary wall formation, a process that includes lignification. As a first step to analyse the corresponding peroxidases, we have isolated previously two anionic isoenzymes (PXP 3-4 and PXP 5) from poplar xylem (Populus trichocarpa), which use SYR as a substrate. Here, we demonstrate that these enzymes are responsible for the visualized SYR oxidation in the developing xylem. The cDNA that corresponds to PXP 3-4 was isolated and the deduced protein was found closely related to the other SYR-oxidizing peroxidase PXP 5 (ca. 98% of identity). PXP 3-4 was expressed in a baculovirus expression system yielding high levels of active peroxidase (3 mg/l medium). The heterologously produced protein showed characteristics similar to those of the corresponding protein from poplar xylem (enzymatic properties, isoelectric point, and migration in a native gel). PXP 3-4 was expressed in the stem and in the root xylem. The data demonstrate that PXP 3-4 (and/or PXP 5) are present in differentiating xylem, supporting a function in secondary cell wall formation.}}, author = {{Christensen, Jørgen Holst and Overney, Serge and Rohde, Antje and Ardiles Diaz, Wilson and Bauw, Guy and Simon, Patrice and Van Montagu, Marc and Boerjan, Wout}}, issn = {{0167-4412}}, journal = {{PLANT MOLECULAR BIOLOGY}}, keywords = {{MOLECULAR-CLONING,SUSPENSION-CULTURES,ANIONIC PEROXIDASE,POPULUS-KITAKAMIENSIS,HORSERADISH-PEROXIDASE,syringaldazine oxidation,secondary cell wall formation,poplar xylem peroxidase,lignification,cDNA isolation,expression analysis,AMINO-ACID,SEQUENCE,LIGNIFICATION,TOBACCO,LIGNIN}}, language = {{eng}}, number = {{5}}, pages = {{581--593}}, title = {{The syringaldazine-oxidizing peroxidase PXP 3-4 from poplar xylem: cDNA isolation, characterization and expression}}, url = {{http://doi.org/10.1023/A:1012271729285}}, volume = {{47}}, year = {{2001}}, }
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