title: Overproduction, purification, and characterization of recombinant aspartate semialdehyde dehydrogenase from Arabidopsis thaliana.
abstract: In plant and microorganisms, aspartate semialdehyde dehydrogenase (ASDH) produces the branch point intermediate between the lysine and threonine/methionine pathways. In this study, we report the first cDNA cloning, purification, and characterization of a plant ASDH. The Arabidopsis thaliana ASDH is an homodimeric enzyme composed of subunits of 36 kDa. The plant enzyme exhibited a specific activity of 26 micromol NADPH oxidized min(-1) mg(-1) of protein with a K(M) value for NADPH of 92 microM. ASDH showed cooperative behavior for aspartyl phosphate with a K(0.5) value of 37 microM.
authors.affiliation: Unité Mixte CNRS/INRA/Aventis CropScience, UMR 1932, 14-20 Rue Pierre Baizet, 69263 Lyon, France.
authors.forename: Stephane
authors.initials: S
authors.lastname: Paris
authors.forename: Peter M
authors.initials: PM
authors.lastname: Wessel
authors.forename: Renaud
authors.initials: R
authors.lastname: Dumas
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D001223:Aspartate-Semialdehyde Dehydrogenase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D007700:Kinetics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D012996:Solutions
chemical_list: D018076:DNA, Complementary; D011994:Recombinant Proteins; D012996:Solutions; D001223:Aspartate-Semialdehyde Dehydrogenase