Rat monoamine oxidase B expressed in Escherichia coli has a covalently-bound FAD.
Rat liver monoamine oxidase B (MAO B) was expressed in E. coli as catalytically active form, though inclusion bodies of the enzyme were also formed as a major protein in the cell. The active form of the recombinant MAO B exhibited similar properties as rat liver enzyme and localized in membrane of the bacteria. Covalent attachment of FAD to polypeptide chain of the recombinant enzyme was revealed by a labeling experiment with [3H]-pargyline, an irreversible mechanism-based inhibitor, indicating that the covalent linkage of FAD to the apoprotein was formed even in the prokaryotic cell. This observation suggests autocatalytic formation of the linkage in MAO B.
Hirashiki
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I
Department of Chemistry, Faculty of Science, Kyushu University, Fukuoka, Japan.
Ogata
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F
Ito
A
A
D000818:Animals; D002462:Cell Membrane / Q000737:chemistry; D005591:Chemical Fractionation; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005182:Flavin-Adenine Dinucleotide / Q000737:chemistry / Q000378:metabolism*; D008099:Liver / Q000201:enzymology; D008995:Monoamine Oxidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010293:Pargyline / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013385:Succinate Dehydrogenase / Q000737:chemistry / Q000378:metabolism
D011994:Recombinant Proteins; D005182:Flavin-Adenine Dinucleotide; D010293:Pargyline; D013385:Succinate Dehydrogenase; D008995:Monoamine Oxidase