title: Purification and characterization of histidinol dehydrogenase from cabbage.
abstract: Histidinol dehydrogenase (EC 1.1.1.23) activity was determined in several plant species and in cultured plant cell lines. The enzyme was purified from cabbage (Brassica oleracea) to apparent homogeneity. To render complete purification, a new, specific histidinol-Sepharose 4B affinity chromatography was developed. The apparent molecular mass of the protein is 103 kDa. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the protein migrated as a single band with a molecular mass of 52 kDa, giving evidence for a dimeric quaternary structure. By isoelectric focusing, the enzyme was separated into six protein bands, five of which possessed the dehydrogenase activity when examined by an activity staining method. The Km values for L-histidinol and NAD+ were 15.5 and 42 microM, respectively. Enzyme activity was stimulated by addition of Mn2+, but was inhibited in the presence of Ba2+, Mg2+, Ni2+, Ca2+, Zn2+, or Cu2+. Histidinol dehydrogenase is the first histidine enzyme that has been purified to homogeneity and characterized from plants. This plant enzyme catalyzes the NAD-linked four-electron dehydrogenase reaction leading from histidinol to His. The results indicate a similar pathway of His in plants and show furthermore the last two reaction steps to be identical to those in microorganisms.
authors.affiliation: International Research Laboratories, CIBA-GEIGY Ltd., Takarazuka, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Nagai
authors.forename: A
authors.initials: A
authors.lastname: Scheidegger
mesh_terms: D000429:Alcohol Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D001937:Brassica / Q000201:enzymology*; D002413:Cations, Divalent / Q000494:pharmacology; D011232:Chemical Precipitation; D002845:Chromatography; D006863:Hydrogen-Ion Concentration; D015394:Molecular Structure; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism
chemical_list: D002413:Cations, Divalent; D009243:NAD; D000429:Alcohol Oxidoreductases; C021358:histidinol dehydrogenase