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  • Beatriz Badia, Mariana (author)

Specific Arabidopsis thaliana malic enzyme isoforms can provide anaplerotic pyruvate carboxylation function in Saccharomyces cerevisiae

  • Article/chapterEnglish2017

Publisher, publication year, extent ...

  • 2017-02-01
  • WILEY,2017
  • printrdacarrier

Numbers

  • LIBRIS-ID:oai:DiVA.org:kth-205137
  • https://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-205137URI
  • https://doi.org/10.1111/febs.14013DOI

Supplementary language notes

  • Language:English
  • Summary in:English

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  • Subject category:ref swepub-contenttype
  • Subject category:art swepub-publicationtype

Notes

  • QC 20170412
  • 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.

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Added entries (persons, corporate bodies, meetings, titles ...)

  • Mans, Robert (author)
  • Lis, Alicia V. (author)
  • Ariel Tronconi, Marcos (author)
  • Lucia Arias, Cintia (author)
  • Maurino, Veronica Graciela (author)
  • Santiago Andreo, Carlos (author)
  • Fabiana Drincovich, Maria (author)
  • van Maris, Antonius J. A.KTH,Industriell bioteknologi(Swepub:kth)u1n330r1 (author)
  • Gerrard Wheeler, Mariel Claudia (author)
  • KTHIndustriell bioteknologi (creator_code:org_t)

Related titles

  • In:The FEBS Journal: WILEY284:4, s. 654-6651742-464X1742-4658

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