Abstract
Red chlorophyll (Chl) catabolite (RCC) reductase, which catalyzes the reaction of an intermediary Chl catabolite (RCC) in the two-step cleavage reaction of pheophorbide (Pheide) a into primary fluorescent catabolites (pFCCs) during Chl breakdown, was characterized and partially purified. RCC reductase activity was present at all stages of barley leaf development and even in roots. The highest specific activity was found in senescent leaves, which were used to purify RCC reductase 1000-fold. Among the remaining three proteins, RCC reductase activity was most likely associated with a 55-kD protein. RCC reductase exhibited saturation kinetics for RCC, with an apparent Michaelis constant of 0.6 mM. The reaction depended on reduced ferredoxin and was sensitive to oxygen. Assays of purified RCC reductase with chemically synthesized RCC as a substrate yielded three different FCCs, two of which could be identified as the stereoisomeric pFCCs from canola (Brassica napus) (pFCC-1) and sweet pepper (Capsicum annuum) (pFCC-2), respectively. In the coupled reaction with Pheide a oxidase and RCC reductase, either pFCC-1 or pFCC-2 was produced, depending on the plant species employed as a source of RCC reductase. Data from 18 species suggest that the stereospecific action of RCC reductase is uniform within a plant family.
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- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Engel N., Jenny T. A., Mooser V., Gossauer A. Chlorophyll catabolism in Chlorella protothecoides. Isolation and structure elucidation of a red bilin derivative. FEBS Lett. 1991 Nov 18;293(1-2):131–133. doi: 10.1016/0014-5793(91)81168-8. [DOI] [PubMed] [Google Scholar]
- Ginsburg S., Matile P. Identification of Catabolites of Chlorophyll-Porphyrin in Senescent Rape Cotyledons. Plant Physiol. 1993 Jun;102(2):521–527. doi: 10.1104/pp.102.2.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ginsburg S., Schellenberg M., Matile P. Cleavage of Chlorophyll-Porphyrin (Requirement for Reduced Ferredoxin and Oxygen). Plant Physiol. 1994 Jun;105(2):545–554. doi: 10.1104/pp.105.2.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matile P., Hortensteiner S., Thomas H., Krautler B. Chlorophyll Breakdown in Senescent Leaves. Plant Physiol. 1996 Dec;112(4):1403–1409. doi: 10.1104/pp.112.4.1403. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morigasaki S., Takata K., Suzuki T., Wada K. Purification and Characterization of a Ferredoxin-NADP Oxidoreductase-Like Enzyme from Radish Root Tissues. Plant Physiol. 1990 Jul;93(3):896–901. doi: 10.1104/pp.93.3.896. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
- Rhie G., Beale S. I. Phycobilin biosynthesis: reductant requirements and product identification for heme oxygenase from Cyanidium caldarium. Arch Biochem Biophys. 1995 Jun 20;320(1):182–194. doi: 10.1006/abbi.1995.1358. [DOI] [PubMed] [Google Scholar]
- Rodoni S., Muhlecker W., Anderl M., Krautler B., Moser D., Thomas H., Matile P., Hortensteiner S. Chlorophyll Breakdown in Senescent Chloroplasts (Cleavage of Pheophorbide a in Two Enzymic Steps). Plant Physiol. 1997 Oct;115(2):669–676. doi: 10.1104/pp.115.2.669. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weller J. L., Terry M. J., Rameau C., Reid J. B., Kendrick R. E. The Phytochrome-Deficient pcd1 Mutant of Pea Is Unable to Convert Heme to Biliverdin IX[alpha]. Plant Cell. 1996 Jan;8(1):55–67. doi: 10.1105/tpc.8.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]