title: Interaction of spin-labeled tryptamine with monoamine oxidase: probing the microenvironment of the active site by spin probe-spin label techniques.
abstract: The spin-labeled substrate, tryptamine, was used as a structural probe of the active site of bovine liver monoamine oxidase B (amine:oxygen oxidoreductase (deaminating) (flavin-containing), EC 1.4.3.4). When the reaction was monitored by electron spin resonance (ESR), line broadening effects indicative of binding with an apparent relation to substrate specificity of the highly purified enzyme were observed. The spectrum indicated that the bound tryptamine was 'partially immobilized' with a dissociation constant of 39 microM and 2.2 mol bound per enzyme dimer. The correlation time, reflecting the environment of the tryptamine binding site, was determined to be 6.2 ns. The topology of the active site was investigated by using dual spin-label methodology in which the spin-labeled substrate, tryptamine, and the 15N-substituted and deuterated maleimide spin label covalently bound to the essential sulfhydryl groups were used. The ESR spectral data suggested that the essential sulfhydryl groups are at least 14 A away from the tryptamine-binding site. The environment surrounding both spin-labeled substrates, tryptamine and [2H,15N]MSL, and the motional properties of the enzyme are discussed.
authors.affiliation: Chemistry Department Atlanta University, GA 30134.
authors.forename: H M
authors.initials: HM
authors.lastname: Zeidan
mesh_terms: D001665:Binding Sites; D004578:Electron Spin Resonance Spectroscopy; D007461:Iodoacetates / Q000494:pharmacology; D019807:Iodoacetic Acid; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008995:Monoamine Oxidase / Q000378:metabolism*; D011487:Protein Conformation; D013113:Spin Labels / Q000378:metabolism*; D014363:Tryptamines / Q000378:metabolism*
chemical_list: D007461:Iodoacetates; D013113:Spin Labels; D014363:Tryptamines; C030820:tryptamine; D008995:Monoamine Oxidase; D019807:Iodoacetic Acid