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Specific Arabidopsis thaliana malic enzyme isoforms can provide anaplerotic pyruvate carboxylation function in Saccharomyces cerevisiae
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Beatriz Badia, Mariana (author)
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Mans, Robert (author)
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Lis, Alicia V. (author)
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Ariel Tronconi, Marcos (author)
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Lucia Arias, Cintia (author)
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Maurino, Veronica Graciela (author)
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Santiago Andreo, Carlos (author)
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Fabiana Drincovich, Maria (author)
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- van Maris, Antonius J. A. (author)
- KTH,Industriell bioteknologi
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Gerrard Wheeler, Mariel Claudia (author)
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(creator_code:org_t)
- 2017-02-01
- 2017
- English.
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In: The FEBS Journal. - : WILEY. - 1742-464X .- 1742-4658. ; 284:4, s. 654-665
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https://febs.onlinel...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Subject headings
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- NAD(P)-malic enzyme (NAD(P)-ME) catalyzes the reversible oxidative decarboxylation of malate to pyruvate, CO2, and NAD(P)H and is present as a multigene family in Arabidopsis thaliana. The carboxylation reaction catalyzed by purified recombinant Arabidopsis NADP-ME proteins is faster than those reported for other animal or plant isoforms. In contrast, no carboxylation activity could be detected in vitro for the NAD-dependent counterparts. In order to further investigate their putative carboxylating role in vivo, Arabidopsis NAD(P)-ME isoforms, as well as the NADP-ME2del2 (with a decreased ability to carboxylate pyruvate) and NADP-ME2R115A (lacking fumarate activation) versions, were functionally expressed in the cytosol of pyruvate carboxylase-negative (Pyc(-)) Saccharomyces cerevisiae strains. The heterologous expression of NADP-ME1, NADP-ME2 (and its mutant proteins), and NADP-ME3 restored the growth of Pyc(-) S. cerevisiae on glucose, and this capacity was dependent on the availability of CO2. On the other hand, NADP-ME4, NAD-ME1, and NAD-ME2 could not rescue the Pyc(-) strains from C-4 auxotrophy. NADP-ME carboxylation activity could be measured in leaf crude extracts of knockout and over-expressing Arabidopsis lines with modified levels of NADP-ME, where this activity was correlated with the amount of NADP-ME2 transcript. These results indicate that specific A. thaliana NADP-ME isoforms are able to play an anaplerotic role in vivo and provide a basis for the study on the carboxylating activity of NADP-ME, which may contribute to the synthesis of C-4 compounds and redox shuttling in plant cells.
Subject headings
- TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology (hsv//eng)
Keyword
- anaplerotic role
- C-4 organic acids
- malate synthesis
- plant metabolism
- Saccharomyces cerevisiae
Publication and Content Type
- ref (subject category)
- art (subject category)
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