Artículo
Specific Arabidopsis thaliana malic enzyme isoforms can provide anaplerotic pyruvate carboxylation function in Saccharomyces cerevisiae
Badia, Mariana Beatriz
; Mans, Robert; Lis, Alicia V.; Tronconi, Marcos Ariel
; Arias, Cintia Lucia
; Maurino, Verónica Graciela; Andreo, Carlos Santiago
; Drincovich, Maria Fabiana
; Maris, Antonius J. A. van; Gerrard Wheeler, Mariel Claudia
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Fecha de publicación:
02/2017
Editorial:
Wiley Blackwell Publishing, Inc
Revista:
Febs Journal
ISSN:
1742-464X
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
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 C4 auxotrophy. NADP-ME carboxylation activity could be measured in leaf crude extracts of knockout and overexpressing 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 C4 compounds and redox shuttling in plant cells.
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Articulos(CEFOBI)
Articulos de CENTRO DE EST.FOTOSINTETICOS Y BIOQUIMICOS (I)
Articulos de CENTRO DE EST.FOTOSINTETICOS Y BIOQUIMICOS (I)
Citación
Badia, Mariana Beatriz; Mans, Robert; Lis, Alicia V.; Tronconi, Marcos Ariel; Arias, Cintia Lucia; et al.; Specific Arabidopsis thaliana malic enzyme isoforms can provide anaplerotic pyruvate carboxylation function in Saccharomyces cerevisiae; Wiley Blackwell Publishing, Inc; Febs Journal; 284; 4; 2-2017; 654-665
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