N-alkanamines as substrates to probe the hydrophobic region of bovine serum amine oxidase active site: A kinetic and spectroscopic study

2007 ◽  
Vol 465 (1) ◽  
pp. 50-60 ◽  
Author(s):  
Maria Luisa Di Paolo ◽  
Carmine Pesce ◽  
Michele Lunelli ◽  
Marina Scarpa ◽  
Adelio Rigo
Keyword(s):  
Spectroscopic Study ◽  
Active Site ◽  
Bovine Serum ◽  
Amine Oxidase ◽  
Hydrophobic Region

scholarly journals Reconstitution of Cu2+-depleted bovine serum amine oxidase with Co2+*

1998 ◽  
Vol 330 (1) ◽  
pp. 383-387 ◽  
Author(s):  
Enzo AGOSTINELLI ◽  
Giovanna DE MATTEIS ◽  
Bruno MONDOVÌ ◽  
Laura MORPURGO
Keyword(s):  
Active Site ◽  
Bovine Serum ◽  
Oxidase Activity ◽  
Amine Oxidase ◽  
Semiquinone Radical ◽  
Copper Site ◽  
Benzylamine Oxidase ◽  
Topa Quinone ◽  
Derivatives Of ◽  
Active Subunits

Two different Cu2+-depleted derivatives of bovine serum amine oxidase (BSAO) have recently been prepared, which contain about 0.5 mol/dimer of phenylhydrazine-reactive topa quinone (TPQ) cofactor and, depending on the reagents used, about 0.2 or 0.7 residual Cu2+/dimer [Agostinelli, De Matteis, Sinibaldi, Mondovì and Morpurgo (1997) Biochem. J. 324, 497-501]. The benzylamine oxidase activity of both derivatives was < 5% and increased up to ≈ 20% on incorporation of Co2+, irrespective of the residual Cu2+ content, which was unaffected by the treatment according to atomic absorption and ESR spectroscopy. The residual Cu2+ ions appeared to be distributed one per dimer and to be bound to inactive subunits, whereas Co2+ was bound to active subunits. The change in the active site had an appreciable influence on the kinetic behaviour. With several amines, the kinetic parameters, Km and kc, measured for Co2+-BSAO were different from those for native BSAO. This excludes the possibility that the catalytic activity was due to residual Cu2+. Furthermore, Co2+ restored to nearly native level the intensity of the TPQ 480 nm band and the reactions with phenylhydrazine or benzylhydrazine, which had been slowed down or abolished, respectively, in Cu2+-depleted samples. The CD spectrum, measured for the derivative with low Cu2+ content, was compatible with Co2+ binding to the copper site. The amine oxidase activity of the Co2+ derivative, which cannot form a semiquinone radical as an intermediate of the catalytic reaction, strongly suggests that the Cu+-semiquinone is not an obligatory intermediate of BSAO catalytic pathway.

scholarly journals Electrostatic compared with hydrophobic interactions between bovine serum amine oxidase and its substrates

2003 ◽  
Vol 371 (2) ◽  
pp. 549-556 ◽  
Author(s):  
Maria Luisa DI PAOLO ◽  
Roberto STEVANATO ◽  
Alessandra CORAZZA ◽  
Fabio VIANELLO ◽  
Lorenzo LUNELLI ◽  
...  
Keyword(s):  
Active Site ◽  
Bovine Serum ◽  
Hydrophobic Interactions ◽  
Amine Oxidase ◽  
Amino Acid Residues ◽  
Deprotonated Form ◽  
Steady State Kinetic ◽  
Wide Range ◽  
Order Of Magnitude

A steady-state kinetic study of bovine serum amine oxidase activity was performed, over a wide range of pH values (5.4–10.2) and ionic strength (10–200mM), using various (physiological and analogue) substrates as specific probes of the active-site binding region. Relatively small changes in kcat values (approx. one order of magnitude) accompanied by marked changes in Km and kcat/Km values (approx. six orders of magnitude) were observed. This behaviour was correlated with the presence of positively charged groups or apolar chains in the substrates. In particular, it was found that the docking of the physiological polyamines, i.e. spermidine and spermine, appears to be modulated by three amino acid residues of the active site, which we have named L-H+, G-H+ and IH+, characterized by pKa values of 6.2±0.2 [Di Paolo, Scarpa, Corazza, Stevanato and Rigo (2002) Biophys. J. 83, 2231–2239], 8.2±0.3 and 7.8±0.4 respectively. The electrostatic interaction between the protonated substrates and the enzyme containing the residues L-H+, G-H+ and IH+ in the deprotonated form, the on/off role of the IH+ residue and the role of hydrophobic interactions with substrates characterized by apolar chains are discussed.

The active site of bovine serum amine oxidase

1983 ◽  
Vol 79 ◽  
pp. 132-133
Author(s):  
Shinnichiro Suzuki ◽  
Takeshi Sakurai ◽  
Akitsugu Nakahara ◽  
Takashi Manabe ◽  
Tsuneo Okuyama
Keyword(s):  
Active Site ◽  
Bovine Serum ◽  
Amine Oxidase

A new redox cofactor in eukaryotic enzymes: 6-hydroxydopa at the active site of bovine serum amine oxidase

Science ◽  
1990 ◽  
Vol 248 (4958) ◽  
pp. 981-987 ◽  
Author(s):  
S. Janes ◽  
D Mu ◽  
D Wemmer ◽  
A. Smith ◽  
S Kaur ◽  
...  
Keyword(s):  
Active Site ◽  
Bovine Serum ◽  
Amine Oxidase

scholarly journals Phosphonium compounds as new and specific inhibitors of bovine serum amine oxidase

2004 ◽  
Vol 384 (3) ◽  
pp. 551-558 ◽  
Author(s):  
Maria Luisa DI PAOLO ◽  
Michele LUNELLI ◽  
Marina SCARPA ◽  
Adelio RIGO
Keyword(s):  
Active Site ◽  
Bovine Serum ◽  
Amine Oxidase ◽  
Cation Binding ◽  
Kinetic Measurements ◽  
New Class ◽  
Molecular Features ◽  
Substrate Structure ◽  
Cation Binding Site ◽  
Competitive Inhibitors

TPP+ (tetraphenylphosphonium ion) and its analogues were found to act as powerful competitive inhibitors of BSAO (bovine serum amine oxidase). The binding of this new class of inhibitors to BSAO was characterized by kinetic measurements. TPP+ can bind to the BSAO active site by hydrophobic and by coulombian interactions. The binding probably occurs in the region of the ‘cation-binding site’[Di Paolo, Scarpa, Corazza, Stevanato and Rigo (2002) Biophys. J. 83, 2231–2239]. Under physiological conditions, the association constant of TPP+ for this site is higher than 106 M−1, the change of enthalpy being the main free-energy term controlling binding. Analysis of the relationships between substrate structure and extent of inhibition by TPP+ reveals some new molecular features of the BSAO active site.

Active site residue involvement in monoamine or diamine oxidation catalysed by pea seedling amine oxidase

FEBS Journal ◽  
2011 ◽  
Vol 278 (8) ◽  
pp. 1232-1243 ◽  
Author(s):  
Maria Luisa Di Paolo ◽  
Michele Lunelli ◽  
Monika Fuxreiter ◽  
Adelio Rigo ◽  
Istvan Simon ◽  
...  
Keyword(s):  
Active Site ◽  
Amine Oxidase ◽  
Active Site Residue ◽  
Pea Seedling

Purification and active-site characterization of equine plasma amine oxidase

1994 ◽  
Vol 56 (2) ◽  
pp. 127-141 ◽  
Author(s):  
Scott R. Carter ◽  
Michele A. McGuirl ◽  
Doreen E. Brown ◽  
David M. Dooley
Keyword(s):  
Active Site ◽  
Amine Oxidase ◽  
Site Characterization ◽  
Equine Plasma ◽  
Plasma Amine Oxidase
Sign in / Sign up

Export Citation Format

Share Document