Maize recombinant non-C4 NADP-malic enzyme: A novel dimeric malic

SAIGO, MARIANA; BOLOGNA, FEDERICO; MAURINO, VERÓNICA GABRIELA; DETARSIO, ENRIQUE; ANDREO, CARLOS; DRINCOVICH, MARÍA FABIANA

PLANT MOLECULAR BIOLOGY

Springer Netherlands

Año: 2004 vol. 55 p. 97 - 107

0167-4412

Among the different isoforms of NADP-malic enzyme (NADP-ME) involved in a wide range of metabolic pathways in plants, the NADP-ME that participates in C4-photosynthesis is the most studied. In the present work, the expression in E. coli of a cDNA encoding for a maize non-photosynthetic NADP-ME is presented. The recombinant NADP-ME thus obtained presents kinetic and structural properties different from the enzyme previously purified from etiolated leaves and roots. Moreover, the recombinant nonphotosynthetic NADP-ME presents very high intrinsic NADP-ME activity, which is unexpected for a non-C4 NADP-ME. Using antibodies against this recombinant enzyme, an immunoreactive band of 66 kDa is detected in different maize tissues indicating that the 66 kDa-NADP-ME is in fact a protein expressedin vivo. The recombinant NADP-ME assembles as a dimer, although the results obtained indicate that ahigher molecular mass oligomeric state of the enzyme is found in maize roots in vivo. In this way, maize presents at least three NADP-ME isoforms: a 72 kDa constitutive form (previously characterized); the novel non-photosynthetic 66 kDa isoform characterized in this work (which is the product of the ZmChlMe2 gene and the likely precursor to the evolution of the photosynthetic C4 NADP-ME) and the 62 kDa isoform (implicated in C4 photosynthesis). The contribution of the present work anticipates further studies concerning the equilibrium between the oligomeric states of the NADP-ME isoforms and the evolution towards the C4 isoenzyme in maize.