Activation of L-lysine epsilon-dehydrogenase from Agrobacterium tumefaciens by several amino acids and monocarboxylates.
The activation of lysine epsilon-dehydrogenase [EC 1.4.1.] by L-lysine was dependent on lysine concentration and was accompanied by association of the dimeric enzymes to a tetramer. The lysine concentration required for the half-maximal activation was 0.28 mM, which was lower than the Km value for L-lysine. In addition to L-lysine, several compounds, which were neither substrates nor inhibitors, activated the enzyme. The compounds which activated the enzyme have common structural characteristics: they have both a carboxyl group and a hydrophobic side chain. These activators also induced the association of the enzyme. The activation of the enzyme occurred well over the pH range 5.0 to 7.5, and the maximal activation was obtained by preincubation for 5 min at 30 degrees C and pH 7.4, when 5 mM L-lysine or 6-aminocaproate was used as an activator. NADH binding experiments indicated that about 2 mol of NADH bind to 1 mol of the tetrameric enzyme: the dimeric enzyme has one catalytic site. Binding experiments with n-[1-14C]heptanoate and L-[U-14C]lysine showed that approximately 2 mol of ligands bind to 1 mol of the dimeric enzyme and L-lysine could not bind to the catalytic site of the enzyme in the absence of NAD+. These results indicate the presence of one catalytic site and two activator binding binding sites in the dimeric enzyme.
Hashimoto
H
H
Department of Agricultural Chemistry, Kochi University, Nankoku.
Misono
H
H
Nagata
S
S
Nagasaki
S
S
D000594:Amino Acid Oxidoreductases / Q000378:metabolism*; D000596:Amino Acids / Q000494:pharmacology*; D001665:Binding Sites; D002264:Carboxylic Acids / Q000494:pharmacology*; D055598:Chemical Phenomena; D002621:Chemistry; D004789:Enzyme Activation; D006863:Hydrogen-Ion Concentration; D009243:NAD / Q000378:metabolism; D012231:Rhizobium / Q000201:enzymology*; D013997:Time Factors
D000596:Amino Acids; D002264:Carboxylic Acids; D009243:NAD; D000594:Amino Acid Oxidoreductases; C034168:lysine epsilon-dehydrogenase