Spectroscopic characterization of carbon monoxide complexes generated for copper/topa quinone-containing amine oxidases.
Carbon monoxide complexes have been generated for copper/topa quinone (TPQ)-containing amine oxidases from Arthrobactor globiformis (AGAO) and Aspergillus niger (AO-I) and characterized by various spectroscopic measurements. Addition of CO to AGAO anaerobically reduced with its substrate 2-phenylethylamine led to a slight increase of absorption bands at 440 and 470 nm derived from the semiquinone form (TPQ(sq)) of the TPQ cofactor, concomitantly giving rise to new CO-related absorption bands at 334 and 434 nm. The intensity of the TPQ(sq) radical EPR signal at g = 2.004 also increased in the presence of CO, while its hyperfine coupling structure was affected insignificantly. FT-IR measurements revealed C-O stretching bands (nu(CO)) at 2063 and 2079 cm(-1) for the CO complex of the substrate-reduced AGAO (at 2085 cm(-1) for AO-I), which shifted nearly 100 cm(-1) to lower frequencies upon using (13)C(18)O. Collectively, these results suggest that CO is bound to the Cu(I) ion in the Cu(I)/TPQ(sq) species formed in the reductive half-reaction of amine oxidation, thereby shifting the Cu(II)/aminoresorcinol right arrow over left arrow Cu(I)/semiquinone equilibrium toward the latter. When AGAO was reduced with dithionite, an intermediary form of the enzyme with Cu(II) reduced to Cu(I) but TPQ still in the oxidized state (TPQ(ox)) was produced. Dithionite reduction of AGAO in the presence of CO resulted in the immediate formation of FT-IR bands at 2064 and 2083 cm(-1), which were assigned to the nu(CO) bands of the CO bound to the TPQ(ox) enzyme. The intense 2083 cm(-1) band was then displaced by a new band at 2077 cm(-1), corresponding to the formation of the fully reduced topa. Significant variation of these nu(CO) frequencies indicates that vibrational properties of CO bound to copper amine oxidases are sensitively influenced by the coordination structure of the Cu(I) ion, which may be modulated by the chemical and redox states of the TPQ cofactor.
Hirota
S
S
Department of Chemistry, Graduate School of Science, Nagoya University, Japan. k46230a@nucc.cc.nagoya-u.ac.jp
Iwamoto
T
T
Tanizawa
K
K
Adachi
O
O
Yamauchi
O
O
D006631:Amine Oxidase (Copper-Containing) / Q000737:chemistry / Q000378:metabolism*; D001173:Arthrobacter / Q000201:enzymology; D001234:Aspergillus niger / Q000201:enzymology; D002248:Carbon Monoxide / Q000737:chemistry / Q000378:metabolism*; D003300:Copper / Q000737:chemistry / Q000378:metabolism; D004295:Dihydroxyphenylalanine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D004227:Dithionite / Q000378:metabolism / Q000494:pharmacology; D004578:Electron Spin Resonance Spectroscopy; D008667:Metalloproteins / Q000737:chemistry / Q000378:metabolism; D010084:Oxidation-Reduction; D013056:Spectrophotometry, Ultraviolet; D017550:Spectroscopy, Fourier Transform Infrared; D013059:Spectrum Analysis, Raman; D013379:Substrate Specificity
D008667:Metalloproteins; D004227:Dithionite; D004295:Dihydroxyphenylalanine; C067626:6-hydroxydopa quinone; D003300:Copper; D002248:Carbon Monoxide; D006631:Amine Oxidase (Copper-Containing)