Comparison of kinetic properties between plant and fungal amine oxidases.
Kinetic properties of novel amine oxidases isolated from a mold Aspergillus niger AKU 3302 were compared to those of typical plant amine oxidase from pea seedling (EC 1.4.3.6). Pea amine oxidase showed highest affinity with diamines, such as putrescine and cadaverine, while fungal enzymes oxidized preferably n-hexylamine and tyramine. All enzymes were inhibited by carbonyl reagents, copper chelating agents, some substrate analogs and alkaloids, but there were quite significant differences in the sensitivity and inhibition modes. Aminoguanidine, which strongly inhibited pea amine oxidases showed only little effect on fungal enzymes. Substrate analogs such as 1.5-diamino-3-pentanone and 1-amino-3-phenyl-3-propanone, which were potent competitive inhibitors of pea amine oxidases, inhibited fungal enzymes much more weakly and non competitively. Also various alkaloids behaving as competitive inhibitors of pea amine oxidase inhibited the fungal enzymes non competitively. Very surprising was the potent inhibition of fungal enzymes by artificial substrates of pea amine oxidases, E- and Z-1,4-diamino-2-butene. The relationships between the different inhibition modes and possible binding at the active site are discussed.
Luhová
L
L
Department of Biochemistry, Faculty of Science, Palacký University, Olomouc, Czech Republic.
Slavík
L
L
Frébort
I
I
Sebela
M
M
Zajoncová
L
L
Pec
P
P
D006631:Amine Oxidase (Copper-Containing); D000588:Amines / Q000378:metabolism*; D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000201:enzymology*; D001665:Binding Sites; D002103:Cadaverine / Q000378:metabolism; D002614:Chelating Agents / Q000494:pharmacology; D003300:Copper / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006146:Guanidines / Q000494:pharmacology; D007659:Ketones / Q000494:pharmacology; D008667:Metalloproteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D008969:Molecular Sequence Data; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D018532:Pisum sativum / Q000201:enzymology*; D010422:Pentanones / Q000494:pharmacology; D011700:Putrescine / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D016415:Sequence Alignment; D013379:Substrate Specificity
D000588:Amines; D002614:Chelating Agents; D004791:Enzyme Inhibitors; D006146:Guanidines; D007659:Ketones; D008667:Metalloproteins; D010422:Pentanones; C021006:diaminobutene; D003300:Copper; C007940:hexylamine; D006631:Amine Oxidase (Copper-Containing); D000587:Oxidoreductases Acting on CH-NH Group Donors; D002103:Cadaverine; C004479:pimagedine; D011700:Putrescine