Development of spin-labeled pargyline analogues as specific inhibitors of human monoamine oxidases A and B.
Three TEMPO-conjugated pargyline analogues (ParSL-1, ParSL-2, and ParSL-3) have been synthesized and their inhibitory properties tested for the two human monoamine oxidase isoforms (hMAOA and hMAOB). The three analogues differ in flexibility and substituent positions (para or meta) of the linkers connecting the TEMPO group to the pargyline phenyl ring. ParSL-1 contains a flexible acetamido (-CH(2)-CO-NH-) linker connecting the two moieties at the para position. In contrast, the TEMPO moieties in ParSL-2 and ParSL-3 are attached with rigid amido (-CO-NH-) linkers to the para or meta positions of the pargyline phenyl ring, respectively. These variations in conformational flexibility and substituent position are shown to have profound effects in tuning the specificities of these analogues toward the two MAO isoforms. ParSL-1 irreversibly inhibits either MAOA and MAOB, ParSL-2 inhibits only MAOB (K(i) = 15 +/- 5 microM), and ParSL-3 is found to be specific for MAOA (K(i) = 268 +/- 72 microM). These results thus provide additional insights into the role of conformational flexibility and structural properties of MAO inhibitors in tuning their isoform specificities. These active site probes have been used to determine the topological orientation of these enzymes in the mitochondrial membrane. Studies with intact mitochondria show MAOA is topologically on the cytosolic face of the outer membrane in human placenta but recombinant MAOA is situated on the opposite inner face in Pichia mitochondria. Recombinant MAOB is found to be situated on the cytosolic face of the outer membrane in Pichia mitochondria.
Upadhyay
Anup K
AK
Department of Biochemistry, Emory University, Atlanta, Georgia 30322, USA.
Edmondson
Dale E
DE
D004578:Electron Spin Resonance Spectroscopy; D006801:Humans; D007700:Kinetics; D008995:Monoamine Oxidase / Q000187:drug effects*; D008996:Monoamine Oxidase Inhibitors / Q000494:pharmacology*; D010293:Pargyline / Q000031:analogs & derivatives* / Q000737:chemistry; D011994:Recombinant Proteins / Q000187:drug effects; D021241:Spectrometry, Mass, Electrospray Ionization; D013056:Spectrophotometry, Ultraviolet; D013113:Spin Labels
D008996:Monoamine Oxidase Inhibitors; D011994:Recombinant Proteins; D013113:Spin Labels; D010293:Pargyline; D008995:Monoamine Oxidase