Amine oxidase from pig plasma (PPAO) has two bound Cu2+ ions and at least one pyrroloquinoline quinone (PQQ) moiety as cofactors. It is shown that recovery of activity by copper-depleted PPAO is linear with respect to added Cu2+ ions. Recovery of e.s.r. and optical spectral characteristics of active-site copper parallel the recovery of catalytic activity. These results are consistent with both Cu2+ ions contributing to catalysis. Further e.s.r. studies indicate that the two copper sites in PPAO, unlike those in amine oxidases from other sources, are chemically distinct. These comparative studies establish that non-identity of the Cu2+ ions in PPAO is not a requirement for amine oxidase activity. It is shown through the use of a new assay procedure that there are two molecules of PQQ bound per molecule of protein in PPAO; only the more reactive of these PQQ moieties is required for activity.
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December 1989
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Research Article|
December 15 1989
Studies on the active site of pig plasma amine oxidase
D Collison;
D Collison
*Department of Chemistry, University of Manchester, Manchester Ml 3 9PL, U.K.
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P F Knowles;
P F Knowles
†Astbury Department of Biophysics, University of Leeds, Leeds LS2 9JT, U.K.
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F E Mabbs;
F E Mabbs
*Department of Chemistry, University of Manchester, Manchester Ml 3 9PL, U.K.
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F X Rius;
F X Rius
†Astbury Department of Biophysics, University of Leeds, Leeds LS2 9JT, U.K.
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I Singh;
I Singh
†Astbury Department of Biophysics, University of Leeds, Leeds LS2 9JT, U.K.
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D M Dooley;
D M Dooley
‡Department of Chemistry, Amherst College, Amherst, MA 01002, U.S.A.
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C E Cote;
C E Cote
‡Department of Chemistry, Amherst College, Amherst, MA 01002, U.S.A.
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M McGuirl
M McGuirl
‡Department of Chemistry, Amherst College, Amherst, MA 01002, U.S.A.
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Publisher: Portland Press Ltd
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 1989 London: The Biochemical Society
1989
Biochem J (1989) 264 (3): 663–669.
Citation
D Collison, P F Knowles, F E Mabbs, F X Rius, I Singh, D M Dooley, C E Cote, M McGuirl; Studies on the active site of pig plasma amine oxidase. Biochem J 15 December 1989; 264 (3): 663–669. doi: https://doi.org/10.1042/bj2640663
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