Inhibition of chickpea seedling copper amine oxidases by tetraethylenepentamine

Document Type: Original article

Authors

1 Deptarment of Biology, Faculty of Science, University of Mohaghegh Ardabili, Ardabil, Iran

2 Deptarment of Science, Faculty of Medicine, University of Medical science, Ardabil, Iran

Abstract

Copper amine oxidases are important enzymes, which contribute to the regulation of mono- and polyamine levels. Each monomer contains one Cu(II) ion and 2,4,5-trihydroxyphenylalanine (TPQ) as cofactors. They catalyze the oxidative deamination of primary amines to aldehydes with a ping-pong mechanism consisting of a transamination. The mechanism is followed by the transfer of two electrons to molecular oxygen which is reduced to hydrogen peroxide. Inhibitors are important tools in the study of catalytic properties of copper amine oxidases and they also have a wide application in physiological research. In this study, purification of the chickpea seedling amine oxidase, was done via salting out by ammonium sulfate and dialysis, followed by DEAE-cellulose column chromatography. By using the Lineweaver - Burk plot, the Km and Vm of the enzyme were found to be 3.3 mM and 0.95 mmol/min/mg, respectively. In this study, the interaction of chickpea diamino oxidase with tetraethylene- pentamine was studied. Analysis of kinetic data indicated that tetraethylenepentamine (with Ki=0.1 mM) inhibits the enzyme by linear mixed inhibitory effect.

Keywords


1. Šebela, M, Tylichová M, Peč P. Inhibition of diamine oxidases and polyamine oxidases by diamine-based compounds. J Neural Transm 2007;114:793–798.

2. Frébort I, Adachi O.Copper/Quinone-Containing Amine Oxidases, an Exciting Class of Ubiquitous Enzymes. J Ferment Bioeng 1995;80:625-632.

3. Longu S, Mura A, Padiglia A, Medda R, Floris G. Mechanism -based inactivators of plant copper/quinone containing amine oxidases. Phytochemistry 2005;66:1751-1758.

4. Heli H, Amani M, Moosavi-Movahedi A, Jabbari A, Floris G, Mura A. Electroactive Centers in Euphorbia Latex and Lentil Seedling Amine Oxidase. Biosci Biotechnol Biochem 2008;72: 29-36.

5. Mura A, Padiglia A, Medda R, Pintus F, Agro A, Floris G. Properties of copper-free pig kidney amine oxidase: Role of topa quinine. FEBS Lett 2006;580:4317–4324.

6. Dawkes H, Phillips S. Copper amine oxidase: cunning cofactor and controversial copper. Curr Opin Struct Biol 2001;11:666–673.

7. Di Paolo M, Vianello F, Stevanato R, Rigo A. Kinetic Characterization of Soybean Seedling Amine Oxidase. Arch Biochem Biophys 1995;323:329-334.

8. Kivirand K, Rinken T. Purification and properties of amine oxidase from pea seedlings. Proc Estonian Acad Sci Chem 2007;56:164–171.

9. Liu YH, Liang WL, Lee CHCH, Tsai YF, Hou WCH. Antioxidant and semicarbazide-sensitive amine oxidase inhibitory activities of glucuronic acid hydroxamate. Food Chem 2011;129: 423–428.

10. Zhang YM, Livingstone JR, Hirasawa E. Purification and characterisation of monoamine oxidase from Avena sativa. Acta Physiol Plant 2012;34:1411–1419.

11. Šebela, M, Radová A, Angelini R, Tavladoraki  P, Frébort I, Peč P. FAD containing polyamine oxidases: a timely challenge for researchers in biochemistry and physiology of plants. Plant Sci 2001;160:197-207.

12. Medda R, Bellelli A, Peč P, Federico R, Cona A, Floris G. Copper amine oxidases of plants. In Copper Amine Oxidases, Floris G, Mondovì B, Ed. Boca Raton: CRC Press, p 2009;44.

13. Story KB. Functional Metabolism: Regulation and Adaptation. Hoboken, NJ: Wiley-Liss 2004.

14. Berg JM, Tymoczko JL, Stryer L. Biochemistry, 5th edition. W. H. Freeman and company 2002.

15. Laemmli UK. Cleavage of structural proteins during the assembly of the bacteriophage T4. Nature 1970;227:680-254.

16. Bardsley WG. Inhibitors of copper amine oxidases. In Structure and Functions of Amine Oxidases, Mondovì B, Ed. Boca Raton: CRC Press, p. 1985;135.

17. Padiglia A, Medda R, Pedersen JZ, Lorrai A, Pec P, Frébort G. Inhibitors of plant copper amine oxidases. J Enzyme Inhibi 1998;13:311-325.

18. Šebela M, Lamplot Z, Petřivalský M, Kopečný D, Lemr K, Frébort I, Peč P. Recent news related to substrates and inhibitors of plant amine oxidases. Biochim Biophys Acta 2003; 1647:355-360.

19. Macholán  L. Substrate-like inhibitors of diamine oxidase: some relations between the structure of aliphatic aminoketones and their inhibitory effect. Arch Biochem Biophys 1969; 134:302-307.

20. Rotilio G. Spectroscopic and chemical properties of the amine oxidase copper. In Structure and Functions of Amine Oxidases, Mondovì B, Ed. Boca Raton: CRC Press, p.1985;127.

21. Devoto G, Massacesi M, Ponticelli G, Medda R, Floris G. Inhibitory activity of bivalent transition-metal complexes with diamines toward a diamine oxidase. Polyhedron 1986;5: 1023-1025.

22. Vianello F, Malek-Mirzayans A, Di Paolo  ML, Stevanato R, Rigo A. Purification and Characterization of Amine Oxidase from Pea Seedlings. Protein Expr Purif 1995;15:196–201.