Investigations of the possible glycosylation of monoamine oxidase B from pig leucocytes.
Monoamine oxidase B (MAO B) from pig liver has been reported to be a sialoglycoprotein. However, when that enzyme from pig lymphocytes and granulocytes was separated by polyacrylamide gel electrophoresis after labelling with the specific irreversible inhibitor [3H]pargyline, staining with 1-ethyl-2-[3-(1-ethyl-naphtho [1,2d] thiazolin-2-ylidene)-2-methylpropenyl] naphtho [1,2d] thiazolium bromide ("Stains-all") failed to detect the presence of sialic acid residues. Treatment of the enzyme in disrupted lymphocytes and granulocytes, or in mitochondrial fractions prepared from them, with neuraminidase resulted in a decrease in MAO activity. However, after the enzyme was rendered soluble by treatment with octylglucoside, treatment with neuraminidase had no effect on the activity. These results indicate that sialic acid residues are not an intrinsic component of MAO B, although associated material containing such groups appears to affect the activity of the membrane-bound enzyme. The activities of membrane-bound preparations of MAO B from pig lymphocytes and granulocytes were unaffected by treatment with trypsin or beta-chymotrypsin. After the preparations had been rendered soluble by treatment with octylglucoside there was a decrease in the activity on treatment with beta-chymotrypsin, but trypsin treatment had no effect. Thus solubilization resulted in residues sensitive to cleavage by the former enzyme becoming accessible to it. Tryptic and chymotryptic peptides separated from the sodium dodecyl sulphate denatured enzymes by polyacrylamide gel electrophoresis revealed no differences between MAO B prepared from lymphocytes and granulocytes.
Balsa
M D
MD
Departament de Bioquímica i Biología Molecular, Facultat de Medicina, Universitat Autónoma de Barcelona, Spain.
Gomez
N
N
Unzeta
M
M
D000818:Animals; D006031:Glycosylation; D006098:Granulocytes / Q000201:enzymology / Q000378:metabolism*; D007962:Leukocytes / Q000201:enzymology / Q000378:metabolism*; D008995:Monoamine Oxidase / Q000302:isolation & purification / Q000378:metabolism*; D009439:Neuraminidase / Q000378:metabolism; D010449:Peptide Mapping; D013552:Swine
D008995:Monoamine Oxidase; D009439:Neuraminidase