SMO (spermine oxidase) and APAO (acetylpolyamine oxidase) are flavoenzymes that play a critical role in the catabolism of polyamines. Polyamines are basic regulators of cell growth and proliferation and their homoeostasis is crucial for cell life since dysregulation of polyamine metabolism has been linked with cancer. In vertebrates SMO specifically catalyses the oxidation of spermine, whereas APAO displays a wider specificity, being able to oxidize both N1-acetylspermine and N1-acetylspermidine, but not spermine. The molecular bases of the different substrate specificity of these two enzymes have remained so far elusive. However, previous molecular modelling, site-directed mutagenesis and biochemical characterization studies of the SMO enzyme–substrate complex have identified Glu216–Ser218 as a putative active site hot spot responsible for SMO substrate specificity. On the basis of these analyses, the SMO double mutants E216L/S218A and E216T/S218A have been produced and characterized by CD spectroscopy and steady-state and rapid kinetics experiments. The results obtained demonstrate that mutation E216L/S218A endows SMO with N1-acetylspermine oxidase activity, uncovering one of the structural determinants that confer the exquisite and exclusive substrate specificity of SMO for spermine. These results provide the theoretical bases for the design of specific inhibitors either for SMO or APAO.
Skip Nav Destination
Article navigation
August 2014
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
- PDF Icon PDF LinkEditorial Board
Research Article|
July 10 2014
The Glu216/Ser218 pocket is a major determinant of spermine oxidase substrate specificity
Manuela Cervelli;
Manuela Cervelli
*Department of Sciences, Roma Tre University, Viale Guglielmo Marconi 446, I-00146 Rome, Italy
†Interuniversity Consortium of Structural and Systems Biology, Viale Medaglie d’Oro 305, I-00136 Rome, Italy
Search for other works by this author on:
Emanuela Angelucci;
Emanuela Angelucci
*Department of Sciences, Roma Tre University, Viale Guglielmo Marconi 446, I-00146 Rome, Italy
Search for other works by this author on:
Pasquale Stano;
Pasquale Stano
*Department of Sciences, Roma Tre University, Viale Guglielmo Marconi 446, I-00146 Rome, Italy
Search for other works by this author on:
Loris Leboffe;
Loris Leboffe
*Department of Sciences, Roma Tre University, Viale Guglielmo Marconi 446, I-00146 Rome, Italy
Search for other works by this author on:
Rodolfo Federico;
Rodolfo Federico
*Department of Sciences, Roma Tre University, Viale Guglielmo Marconi 446, I-00146 Rome, Italy
Search for other works by this author on:
Giovanni Antonini;
Giovanni Antonini
*Department of Sciences, Roma Tre University, Viale Guglielmo Marconi 446, I-00146 Rome, Italy
†Interuniversity Consortium of Structural and Systems Biology, Viale Medaglie d’Oro 305, I-00136 Rome, Italy
Search for other works by this author on:
Paolo Mariottini;
Paolo Mariottini
*Department of Sciences, Roma Tre University, Viale Guglielmo Marconi 446, I-00146 Rome, Italy
†Interuniversity Consortium of Structural and Systems Biology, Viale Medaglie d’Oro 305, I-00136 Rome, Italy
Search for other works by this author on:
Fabio Polticelli
Fabio Polticelli
1
*Department of Sciences, Roma Tre University, Viale Guglielmo Marconi 446, I-00146 Rome, Italy
‡National Institute of Nuclear Physics, Roma Tre Section, Via della vasca navale 84, I-00146 Rome, Italy
1To whom correspondence should be addressed (email polticel@uniroma3.it).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
March 06 2014
Revision Received:
May 06 2014
Accepted:
May 23 2014
Accepted Manuscript online:
May 23 2014
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2014 Biochemical Society
2014
Biochem J (2014) 461 (3): 453–459.
Article history
Received:
March 06 2014
Revision Received:
May 06 2014
Accepted:
May 23 2014
Accepted Manuscript online:
May 23 2014
Citation
Manuela Cervelli, Emanuela Angelucci, Pasquale Stano, Loris Leboffe, Rodolfo Federico, Giovanni Antonini, Paolo Mariottini, Fabio Polticelli; The Glu216/Ser218 pocket is a major determinant of spermine oxidase substrate specificity. Biochem J 1 August 2014; 461 (3): 453–459. doi: https://doi.org/10.1042/BJ20140305
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.