Amino acid sequence similarity between malate dehydrogenases (NAD) and pea chloroplast malate dehydrogenase (NADP).
Purified pea chloroplast malate dehydrogenase (NADP) was reduced, S-pyridylethylated with 4-vinyl-pyridine and cleaved with trypsin. The resulting peptides were separated by reversed-phase high-performance liquid chromatography. Several of these peptides were subjected to automated Edman degradation. The sequences obtained were compared to the published primary structures of malate dehydrogenase from the thermophilic bacterium Thermus flavus and with the sequence of heart mitochondrial and cytoplasmic malate dehydrogenase (NAD). Most peptides from choroplast malate dehydrogenase (NADP) showed high homology with sequences of the other malate dehydrogenases, especially with those of the bacterial enzyme. One of the sequenced peptides contains the active-site histidine residue which is conserved in all malate dehydrogenases. Our results suggest a common evolutionary origin for all malate dehydrogenases despite their different coenzyme specificities and regulatory properties. The sequenced peptides which revealed no homology were either located at the amino-terminal or at the carboxy-terminal region of chloroplast malate dehydrogenase (NADP). These novel sequences are most likely plant-specific extensions of an ancestral malate dehydrogenase and may be responsible for the unique light-dependent activation of the chloroplast enzyme.
Fickenscher
K
K
Department of Biological Chemistry and Structure, University of Health Sciences, Chicago Medical School, Illinois.
Scheibe
R
R
Marcus
F
F
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002736:Chloroplasts / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid; D003600:Cytosol / Q000201:enzymology; D007887:Fabaceae / Q000201:enzymology; D006868:Hydrolysis; D008291:Malate Dehydrogenase / Q000032:analysis*; D008929:Mitochondria, Heart / Q000201:enzymology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D010454:Peptide Termination Factors / Q000032:analysis; D010946:Plants, Medicinal; D013552:Swine; D013824:Thermus / Q000201:enzymology; D014357:Trypsin
D000596:Amino Acids; D010446:Peptide Fragments; D010454:Peptide Termination Factors; D008291:Malate Dehydrogenase; C046446:malate dehydrogenase-(oxaloacetate-decarboxylating) (NAD+); C031917:malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+); D014357:Trypsin