Chloroplast Biogenesis 72: A [4-Vinyl]Chlorophyllide a Reductase Assay Using Divinyl Chlorophyllide a as an Exogenous Substrate

Parham, Ramin; Rebeiz, Constantin A.

doi:10.1006/abio.1995.1516

Cited by 40 publications

(25 citation statements)

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“…Using this system, they showed that DVR is able to reduce the 8-vinyl group of DV-Chlide a, but it cannot react with 3,8-DVPchlide or DV-Chlide b at observable rates in vitro. By using an in vivo method with isolated cucumber etioplast membranes, Parham and Rebeiz (1995) also report that DV-Chlide a can be readily converted to MV-Chlide. The rate of this conversion is estimated to be 50-to 300-fold higher than that of the activity to convert DV-Pchlide to MV-Pchlide in isolated barley etioplasts (Tripathy and Rebeiz 1988).…”

Section: 8-divinyl Pchlide a 8-vinyl Reductasementioning

confidence: 90%

Tetrapyrrole Metabolism inArabidopsis thaliana

Tanaka

Kobayashi²,

Masuda

2011

The Arabidopsis Book

229

232

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This manuscript is dedicated to Prof. Mamoru Mimuro of Kyoto University who passed away in Februay 2011.Higher plants produce four classes of tetrapyrroles, namely, chlorophyll (Chl), heme, siroheme, and phytochromobilin. In plants, tetrapyrroles play essential roles in a wide range of biological activities including photosynthesis, respiration and the assimilation of nitrogen/sulfur. All four classes of tetrapyrroles are derived from a common biosynthetic pathway that resides in the plastid. In this article, we present an overview of tetrapyrrole metabolism in Arabidopsis and other higher plants, and we describe all identified enzymatic steps involved in this metabolism. We also summarize recent findings on Chl biosynthesis and Chl breakdown. Recent advances in this field, in particular those on the genetic and biochemical analyses of novel enzymes, prompted us to redraw the tetrapyrrole metabolic pathways. In addition, we also summarize our current understanding on the regulatory mechanisms governing tetrapyrrole metabolism. The interactions of tetrapyrrole biosynthesis and other cellular processes including the plastid-to-nucleus signal transduction are discussed.

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Section: 8-divinyl Pchlide a 8-vinyl Reductasementioning

confidence: 90%

Tetrapyrrole Metabolism inArabidopsis thaliana

Tanaka

Kobayashi²,

Masuda

2011

The Arabidopsis Book

229

232

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“…This activity has been detected with isolated plastid membranes (Parham and Rebeiz, 1995) from cucumber (Cucumis sativus) and in solubilized crude extracts (Kolossov and Rebeiz, 2001). It has been proposed that 8-vinyl group reduction may occur at various steps of chlorophyll biosynthesis before and after the reduction of D pyrrole ring by protochlorophyllide oxidoreductase.…”

Section: Introductionmentioning

confidence: 97%

“…DVR performs reduction of 8-vinyl group on the tetrapyrrole to an ethyl group (Parham and Rebeiz, 1995) using NADPH as the reductant. This activity has been detected with isolated plastid membranes (Parham and Rebeiz, 1995) from cucumber (Cucumis sativus) and in solubilized crude extracts (Kolossov and Rebeiz, 2001).…”

Section: Introductionmentioning

confidence: 99%

“…It has been proposed that 8-vinyl group reduction may occur at various steps of chlorophyll biosynthesis before and after the reduction of D pyrrole ring by protochlorophyllide oxidoreductase. As a result, it is likely that the monovinyl and divinyl chlorophyll biosynthesis reactions may operate in parallel rather than by the normally accepted single, linear pathway (Parham and Rebeiz, 1995). However, at this time, it is unclear whether different enzymes or a single enzyme are responsible for the reduction of the 8-vinyl group of various intermediate molecules of chlorophyll biosynthesis.…”

Section: Introductionmentioning

confidence: 99%

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Identification of a Vinyl Reductase Gene for Chlorophyll Synthesis in Arabidopsis thaliana and Implications for the Evolution of Prochlorococcus Species

et al. 2005

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Chlorophyll metabolism has been extensively studied with various organisms, and almost all of the chlorophyll biosynthetic genes have been identified in higher plants. However, only the gene for 3,8-divinyl protochlorophyllide a 8-vinyl reductase (DVR), which is indispensable for monovinyl chlorophyll synthesis, has not been identified yet. In this study, we isolated an Arabidopsis thaliana mutant that accumulated divinyl chlorophyll instead of monovinyl chlorophyll by ethyl methanesulfonate mutagenesis. Map-based cloning of this mutant resulted in the identification of a gene (AT5G18660) that shows sequence similarity with isoflavone reductase genes. The mutant phenotype was complemented by the transformation with the wild-type gene. A recombinant protein encoded by AT5G18660 was expressed in Escherichia coli and found to catalyze the conversion of divinyl chlorophyllide to monovinyl chlorophyllide, thereby demonstrating that the gene encodes a functional DVR. DVR is encoded by a single copy gene in the A. thaliana genome. With the identification of DVR, finally all genes required for chlorophyll biosynthesis have been identified in higher plants. Analysis of the complete genome of A. thaliana showed that it has 15 enzymes encoded by 27 genes for chlorophyll biosynthesis from glutamyl-tRNAglu to chlorophyll b. Furthermore, identification of the DVR gene helped understanding the evolution of Prochlorococcus marinus, a marine cyanobacterium that is dominant in the open ocean and is uncommon in using divinyl chlorophylls. A DVR homolog was not found in the genome of P. marinus but found in the Synechococcus sp WH8102 genome, which is consistent with the distribution of divinyl chlorophyll in marine cyanobacteria of the genera Prochlorococcus and Synechococcus

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“…Varieties of reports lead to the assumption that Chl biosynthetic heterogeneity originates mainly in parallel DV-and MVChl biosynthetic routes. These routes are interconnected by 8-vinyl reductases that convert DV-tetrapyrroles to MV-tetrapyrroles by conversion of the vinyl group at position 8 of ring B to the ethyl group (Parham and Rebeiz, 1995;Rebeiz et al, 2003). DV-MPE could be converted to MV-MPE in crude homogenates from etiolated wheat (Triticum aestivum) seedlings (Ellsworth and Hsing, 1974).…”

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confidence: 99%

One Divinyl Reductase Reduces the 8-Vinyl Groups in Various Intermediates of Chlorophyll Biosynthesis in a Given Higher Plant Species, But the Isozyme Differs between Species

Wang

Wan

et al. 2012

Plant Physiology

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Divinyl reductase (DVR) converts 8-vinyl groups on various chlorophyll intermediates to ethyl groups, which is indispensable for chlorophyll biosynthesis. To date, five DVR activities have been detected, but adequate evidence of enzymatic assays using purified or recombinant DVR proteins has not been demonstrated, and it is unclear whether one or multiple enzymes catalyze these activities. In this study, we systematically carried out enzymatic assays using four recombinant DVR proteins and five divinyl substrates and then investigated the in vivo accumulation of various chlorophyll intermediates in rice (Oryza sativa), maize (Zea mays), and cucumber (Cucumis sativus). The results demonstrated that both rice and maize DVR proteins can convert all of the five divinyl substrates to corresponding monovinyl compounds, while both cucumber and Arabidopsis (Arabidopsis thaliana) DVR proteins can convert three of them. Meanwhile, the OsDVR (Os03g22780)-inactivated 824ys mutant of rice exclusively accumulated divinyl chlorophylls in its various organs during different developmental stages. Collectively, we conclude that a single DVR with broad substrate specificity is responsible for reducing the 8-vinyl groups of various chlorophyll intermediates in higher plants, but DVR proteins from different species have diverse and differing substrate preferences, although they are homologous.

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Chloroplast Biogenesis 72: A [4-Vinyl]Chlorophyllide a Reductase Assay Using Divinyl Chlorophyllide a as an Exogenous Substrate

Cited by 40 publications

References 0 publications

Tetrapyrrole Metabolism inArabidopsis thaliana

Tetrapyrrole Metabolism inArabidopsis thaliana

Identification of a Vinyl Reductase Gene for Chlorophyll Synthesis in Arabidopsis thaliana and Implications for the Evolution of Prochlorococcus Species

One Divinyl Reductase Reduces the 8-Vinyl Groups in Various Intermediates of Chlorophyll Biosynthesis in a Given Higher Plant Species, But the Isozyme Differs between Species

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