Characterization of monoamine oxidase activity present in human granulocytes and lymphocytes.
The characterization of monoamine oxidase (MAO) activity in lymphocytes and granulocytes was studied by using cells prepared from human blood. The specific activities of the enzyme towards beta-phenylethylamine (PEA), benzylamine (Bz), tyramine (TYR) and 5-hydroxytryptamine (5-HT) were found to be 5-times higher in lymphocytes than in granulocytes. The absence of the semicarbazide-sensitive amine oxidase (SSAO) was confirmed by the lack of effect of semicarbazide on the benzylamine oxidation. The presence of MAO-B was corroborated by the inhibition of PEA oxidation with nanomolar deprenyl concentrations and by inhibition of TYR oxidation with high clorgyline concentrations, as well as by the simple sigmoid curve obtained in both cases. These results, together with the substrate preferences, suggest that the MAO activity of human granulocytes and lymphocytes is predominantly of the B form. For each fraction the kinetic constants were determined towards PEA, TYR and Bz as substrates. The Km values were similar for both cellular samples, whereas the Vmax values were higher in lymphocytes than in granulocytes. MAO-B was titrated with [3H]pargyline in order to find out the number of active sites. The corresponding molecular concentration, Kcat values and turnover number showed the presence of related enzymes in human granulocytes and lymphocytes.
Balsa
M D
MD
Departament de Bioquimica i Biologia Molecular, Facultat de Medicina, Universitat Autònoma de Barcelona, Spain.
Gómez
N
N
Unzeta
M
M
D000328:Adult; D001665:Binding Sites; D003010:Clorgyline / Q000494:pharmacology; D006098:Granulocytes / Q000201:enzymology* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008214:Lymphocytes / Q000201:enzymology* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D008995:Monoamine Oxidase / Q000097:blood* / Q000378:metabolism; D008996:Monoamine Oxidase Inhibitors / Q000494:pharmacology; D010293:Pargyline / Q000378:metabolism; D012642:Selegiline / Q000494:pharmacology; D012663:Semicarbazides / Q000494:pharmacology; D013379:Substrate Specificity
D008996:Monoamine Oxidase Inhibitors; D012663:Semicarbazides; D012642:Selegiline; C010059:carbamylhydrazine; D010293:Pargyline; D008995:Monoamine Oxidase; D003010:Clorgyline