Identification of Fructose-1,6-bisphosphate aldolase cytosolic class I as an NMH7 MADS domain associated protein.
We are interested in identifying proteins that interact with the MADS domain protein NMH7 of Medicago sativa. We use an affinity column with a synthetic peptide derived from the MADS domain of NMH7 which has been reported to mediate protein-protein interaction with non-MADS domain interacting proteins. We identified approximately 40 and approximately 80kDa specifically bound proteins as the monomeric and dimeric forms of Fructose-1,6-bisphosphate aldolase cytosolic class I. NiNTA pull down assays revealed that K- and C-terminus regions of NMH7 are not required for the interaction with aldolase. Aldolase enzymatic activity is not required for the interaction with NMH7. NMH7 and aldolase were coimmunoprecipitated from non-inoculated seed and seedlings extracts. Colocalization studies using confocal microscopy showed that aldolase and NMH7 are localized in the cytoplasm and the nucleus of the cortical cells. These data together show that M. sativa aldolase is a novel MADS domain binding protein, and suggest a broader functional repertory for this enzyme, as has been proposed for other glycolytic enzymes.
Páez-Valencia
Julio
J
Departamento de Ecología Funcional, Instituto de Ecología, Universidad Nacional Autónoma de México, Ciudad Universitaria 04510, México DF, Mexico.
Valencia-Mayoral
Pedro
P
Sánchez-Gómez
Concepción
C
Contreras-Ramos
Alejandra
A
Hernández-Lucas
Ismael
I
Martínez-Barajas
Eleazar
E
Gamboa-DeBuen
Alicia
A
D000595:Amino Acid Sequence; D002467:Cell Nucleus / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism*; D006019:Glycolysis; D026161:MADS Domain Proteins / Q000378:metabolism*; D000455:Medicago sativa / Q000201:enzymology*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000378:metabolism*; D036226:Seedlings / Q000201:enzymology; D012639:Seeds / Q000201:enzymology
D026161:MADS Domain Proteins; D010940:Plant Proteins; D005634:Fructose-Bisphosphate Aldolase