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.
Skip Nav Destination
Article navigation
April 2003
- PDF Icon PDF LinkFront Matter
Research Article|
April 15 2003
Electrostatic compared with hydrophobic interactions between bovine serum amine oxidase and its substrates
Maria Luisa DI PAOLO;
Maria Luisa DI PAOLO
∗Dipartimento di Chimica Biologica, Università di Padova, Via G. Colombo 3, 35121 Padova, Italy
Search for other works by this author on:
Roberto STEVANATO;
Roberto STEVANATO
†Dipartimento di Chimica Fisica, Università di Venezia, 30100 Venezia, Italy
Search for other works by this author on:
Alessandra CORAZZA;
Alessandra CORAZZA
‡Dipartimento di Scienze e Tecnologie Biomediche, Università di Udine, 33100 Udine, Italy
Search for other works by this author on:
Fabio VIANELLO;
Fabio VIANELLO
∗Dipartimento di Chimica Biologica, Università di Padova, Via G. Colombo 3, 35121 Padova, Italy
Search for other works by this author on:
Lorenzo LUNELLI;
Lorenzo LUNELLI
§Dipartimento di Fisica and INFM, Università di Trento, Via Sommarive 14, 38050 Povo-Trento, Italy
Search for other works by this author on:
Marina SCARPA;
Marina SCARPA
§Dipartimento di Fisica and INFM, Università di Trento, Via Sommarive 14, 38050 Povo-Trento, Italy
Search for other works by this author on:
Adelio RIGO
Adelio RIGO
1
∗Dipartimento di Chimica Biologica, Università di Padova, Via G. Colombo 3, 35121 Padova, Italy
1To whom correspondence should be addressed (e-mail adelio.rigo@unipd.it).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
July 04 2002
Revision Received:
November 06 2002
Accepted:
January 15 2003
Accepted Manuscript online:
January 15 2003
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2003
2003
Biochem J (2003) 371 (2): 549–556.
Article history
Received:
July 04 2002
Revision Received:
November 06 2002
Accepted:
January 15 2003
Accepted Manuscript online:
January 15 2003
Citation
Maria Luisa DI PAOLO, Roberto STEVANATO, Alessandra CORAZZA, Fabio VIANELLO, Lorenzo LUNELLI, Marina SCARPA, Adelio RIGO; Electrostatic compared with hydrophobic interactions between bovine serum amine oxidase and its substrates. Biochem J 15 April 2003; 371 (2): 549–556. doi: https://doi.org/10.1042/bj20021055
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.