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2006 | 53 | 3 | 539-546
Article title

Influence of dipeptidyl peptidase IV on enzymatic properties of adenosine deaminase

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The importance of ADA (adenosine deaminase) in the immune system and the role of its interaction with an ADA-binding cell membrane protein dipeptidyl peptidase IV (DPPIV), identical to the activated immune cell antigen, CD26, has attracted the interest of researchers for many years. To investigate the specific properties in the structure - function relationship of the ADA/DPPIV-CD26 complex, its soluble form, identical to large ADA (LADA), was isolated from human blood serum, human pleural fluid and bovine kidney cortex. The kinetic constants (Km and Vmax) of LADA and of small ADA (SADA), purified from bovine lung and spleen, were compared using adenosine (Ado) and 2'-deoxyadenosine (2'-dAdo) as substrates. The Michaelis constant, Km, evidences a higher affinity of both substrates (in particular of more toxic 2'-dAdo) for LADA and proves the modulation of toxic nucleoside neutralization in the extracellular medium due to complex formation between ADA and DPPIV-CD26. The values of Vmax are significantly higher for SADA, but the efficiency, Vmax /Km, in LADA-catalyzed 2'-dAdo deamination is higher than that in Ado deamination. The interaction of all enzyme preparations with derivatives of adenosine and erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) was studied. 1-DeazaEHNA and 3-deazaEHNA demonstrate stronger inhibiting activity towards LADA, the DPPIV-CD26-bound form of ADA. The observed differences between the properties of the two ADA isoforms may be considered as a consequence of SADA binding with DPPIV-CD26. Both SADA and LADA indicated a similar pH-profile of adenosine deamination reaction with the optimum at pHs 6.5 - 7.5, while the pH-profile of dipeptidyl peptidase activity of the ADA/DPPIV-CD26 complex appeared in a more alkaline region.
Physical description
  • H.Ch. Buniatyan Institute of Biochemistry, Yerevan, Armenia
  • H.Ch. Buniatyan Institute of Biochemistry, Yerevan, Armenia
  • H.Ch. Buniatyan Institute of Biochemistry, Yerevan, Armenia
  • Department of Biology M.C.A. Sciences, University of Camerino, Camerino, Italy
  • Department of Chemical Sciences, University of Camerino, Camerino, Italy
  • Akedo H, Nishihara H, Shinkai K, Komatsu K, Ishikawa S (1972) Multiple forms of human adenosine deaminase. I Purification and characterization of two molecular species. Biochim Biophys Acta 276: 257-271.
  • Beraudi A, Traversa U, Villani L, Sekino Y, Nagy JI, Poli A (2003) Distribution and expression of A1 adenosine receptors, adenosine deaminase and adenosine deaminase-binding protein (CD26) in goldfish brain. Neurochem Int 42: 455-464.
  • Boonacker CJ, Van Noorden CJF (2003) The moonlighting or multifunctional protein CD26/DPPIV. Eur J Cell Biol 82: 53-73.
  • Bradford MM (1976) A rapid and sensitive method for quantization of microgram quantities of protein utilizing the principal of protein-dye binding. Anal Biochem 72: 248-254.
  • Cristalli G, Eleuteri A, Vittori S, Volpini R, Camaioni E, Lupidi G (1993) Adenosine deaminase inhibitors - structure-activity relationships in 1-deasaadenosine and erithro-9-(2-hydroxy-3-nonyl)adenine analogs. Drug Dev Res 28: 253-258.
  • Daddona PE, Kelley WN (1978) Human adenosine deaminase binding protein. Assay, purification and properties. J Biol Chem 253: 4617-4623.
  • De Meester I, Vanham G, Kestens L, Vanhoof G, Bosmans E, Gigase P, Scharpe S (1994) Binding of adenosine deaminase to the lymphocyte surface via CD26. Eur J Immunol 24: 566-570.
  • Dong RP, Kameoka J, Hegen M, Tanaka T, Xu Y, Schlossman SF, Morimoto C (1996) Characterization of adenosine binding to human CD26 on T cells and its biologic role in immune response. J Immunol 156: 1349-1355.
  • Fonoll C, Canela EI, Bozal J (1982) Characterization of the forms of bovine liver adenosine deaminase. Int J Biochem 14: 679-683.
  • Fox DA, Hussey RE, Fitzgerald KA, Acuto O, Poole C, Palley L, Daley JF, Schlossman SF, Reinherz EL (1984) Ta1, a novel 105 KD human T cell activation antigen defined by monoclonal antibody. J Immunol 133: 1250-1256.
  • Franco R, Mallol J, Casado V, Lluis C, Canela EI, Saura C, Blanco J, Ciruela F (1998) Ecto-adenosine deaminase: an ecto-enzyme and a co-stimulatory protein acting on a variety of cell surface receptors. Drug Dev Res 45: 261-268.
  • Frieden C, Kurz LC, Gilbert HR (1980) Adenosine deaminase and adenylate deaminase: comparative kinetic studies with transition state and ground state analogue inhibitors. Biochemistry 19: 5303-5309.
  • Giblett ER, Anderson JE, Cohen F, Pollara B, Meuwissen HJ (1972) Adenosine deaminase deficiency in two patients with severely impaired cellular immunity. Lancet 2: 1067-1069.
  • Gines S, Ciruela F, Burgueno J, Casado V, Canela EI, Mallol J, Lluis C, Franco R (2001) Involvement of caveoline in ligand-induced recruitment and internalization of A1 adenosine receptor and adenosine deaminase in an epithelial cell line. Mol Pharmacol 59: 1314-1323.
  • Gorrell MD (2005) Dipeptidyl peptidase IV and related enzymes in cell biology and liver disorders. Clin Sci 108: 277-292.
  • Gorrell MD, Gysbers V, McCaughan GW (2001) CD26: a multifunctional integral membrane and secreted protein of activated lymphocytes. Scand J Immunol 54: 249-264.
  • Hirschhorn R (1990) Adenosine deaminase deficiency. Immunodefic Rev 2: 175-198.
  • Kameoka J, Tanaka T, Nojima Y, Schlossman S, Morimoto C (1993) Direct association of adenosine deaminase with T cell activation antigen, CD26. Science 261: 466-469.
  • Leatherbarrow RJ (2001) GraFit Version 5, Erithacus Software Ltd., Horley, U.K.
  • Ludwig K, Fan H, Dobers J, Berger M, Reutter W, Bottcher C (2004) 3D structure of the CD26-ADA complex obtained by cryo-EM and single particle analysis. Biochem Biophys Res Comm 313: 223-229.
  • Mardanyan S, Sharoyan S, Antonyan A, Armenyan A, Cristalli G, Lupidi G (2001) Tryptophan environment in adenosine deaminase: enzyme modification with N-bromosuccinimide in the presence of adenosine and EHNA analogs. Biochim Biophys Acta 1546: 185-195.
  • Martin M, Huguet J, Centelles JJ, Franco R (1995) Expression of ecto-adenosine deaminase and CD26 in human T cells triggered by TCR-CD3 complex: possible role of adenosine deaminase as costimulatory molecule. J Immunol 155: 4630-4643.
  • Mentlein R (2004) Cell surface peptidases. Int Rev Cytol 235: 165-213.
  • Mentlein R, Struckhoff G (1989) Purification of two dipeptidyl aminopeptidase II from rat brain and their action on proline-containing neuropeptides. J Neurochem 52: 1284-1293.
  • Murphy JB, Kies MW (1960) Note on spectrophotometric determination of proteins in dilute solutions. Biochim Biophys Acta 45: 382-384.
  • Schrader WP, Stacy AR (1977) Purification and subunit structure of adenosine deaminase from human kidney. J Biol Chem 252: 6409-6416.
  • Senba E, Daddona PE, Nagy I (1987) A subpopulation of preganglionic parasympathetic neurons in the rat contain adenosine deaminase. Neuroscience 20: 487-502.
  • Sharoyan SG, Antonyan AA, Mardanyan SS (1994) Isolation, purification and comparative investigation of adenosine deaminase from five parts of cattle brain. Biochemistry (Moscow) 59: 169-174.
  • Trotta PP (1982) Identification of a membrane adenosine deaminase binding protein from human plasma. Biochemistry 21: 4014-4023.
  • Trotta PP, Peterfreund RA, Schonberg R, Balis ME (1979) Rabbit adenosine deaminase conversion proteins. Purification and crystallization. Biochemistry 18: 2953-2958.
  • Van der Weyden MB, Kelley WN (1976) Human adenosine deaminase. Distribution and properties. J Biol Chem 251: 5448-5456.
  • Van der Weyden MB, Kelley WN (1977) Adenosine deaminase deficiency and severe combined immunodeficiency disease. Life Sciences 20: 1645-1650.
  • Weisman MI, Caiolfa VR, Parola AH (1988) Adenosine deaminase-complexing protein from bovine kidney. Isolation of two distinct subunits. J Biol Chem 263: 5266-5270.
  • Wilson DK, Rudolf FB, Quiocho FA (1991) Atomic structure of adenosine deaminase complexed with a transition-state analog: understanding catalysis and immunodeficiency mutations. Science 252: 1278-1284.
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