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Abstract:

Cyclic nucleotide phosphodiesterases constitute the only known mechanism to inactivate regulatory signals involving cAMP or cGMP. In our laboratory a cAMP-specific phosphodiesterase associated to the flagellar apparatus, named TcPDE1, was identified in Trypanosoma cruzi. By using the catalytic domain sequence of TcPDE1 to screen a Trypanosoma cruzi genomic data base, a novel T. cruzi phosphodiesterase sequence was found and characterized. TcPDE4 encodes a 924-amino acid protein and shows homology with the PDE4 vertebrate subfamily. The sequence shows three conserved domains, FYVE, phosphohydrolase and PDEaseI. The FYVE zinc-finger domain is characteristic of proteins recruited to phosphatidylinosytol 3-phosphate-containing membranes, whereas the two others are characteristic of phosphohydrolases and members of the cyclic nucleotide phosphodiesterases. Sequence analysis shows all characteristic domains present at the type-4 phosphodiesterases specific for cAMP. Moreover, TcPDE4 shows the inhibition profile characteristic for PDE4 subfamily, with an IC50 of 10.46 μM for rolipram and 1.3 μM for etazolate. TcPDE4 is able to complement a heat-shock-sensitive yeast mutant deficient in phosphodiesterase genes. The enzyme is specific for cAMP, Mg2+-dependent and its activity is not affected by cGMP or Ca2+. The association of TcPDE4 with membranes was studied by subcellular fractionation of recombinant yeast and extraction in several conditions. Most of the enzyme remained associated to the membrane fraction after treatment with high salt concentration, detergent, or chaotropic agents. This support previous hypotheses that in this parasite cAMP phosphodiesterases, and consequently cAMP levels, are compartimentalized. © 2005 Elsevier B.V. All rights reserved.

Registro:

Documento: Artículo
Título:TcPDE4, a novel membrane-associated cAMP-specific phosphodiesterase from Trypanosoma cruzi
Autor:Alonso, G.D.; Schoijet, A.C.; Torres, H.N.; Flawiá, M.M.
Filiación:Instituto de Investigaciones en Ingeniería Genética Y Biología Molecular, Consejo Nacional de Investigaciones Científicas Y Técnicas, Universidad de Buenos Aires, Vuelta de Obligado 2490, 1428 Buenos Aires, Argentina
Palabras clave:cAMP; FYVE domain; Phosphatidylinositol 3-phosphate; Rolipram; detergent; etazolate; membrane protein; phosphatase; phosphatidylinositol 3 phosphate; phosphodiesterase IV; protozoal protein; rolipram; sodium chloride; amino acid sequence; article; cell compartmentalization; cell membrane; concentration (parameters); enzyme active site; enzyme inhibition; enzyme specificity; eukaryotic flagellum; extraction; gene; genetic conservation; genome analysis; IC 50; nonhuman; nucleotide sequence; priority journal; protein analysis; protein family; screening; sequence analysis; sequence homology; temperature sensitive mutant; Trypanosoma cruzi; vertebrate; zinc finger motif; 3',5'-Cyclic-Nucleotide Phosphodiesterase; Amino Acid Sequence; Animals; Base Sequence; Cell Membrane; Cloning, Molecular; DNA, Protozoan; Etazolate; Kinetics; Molecular Sequence Data; Phosphodiesterase Inhibitors; Recombinant Proteins; Rolipram; Sequence Alignment; Trypanosoma cruzi; Eukaryota; Trypanosoma cruzi; Vertebrata
Año:2006
Volumen:145
Número:1
Página de inicio:40
Página de fin:49
DOI: http://dx.doi.org/10.1016/j.molbiopara.2005.09.005
Título revista:Molecular and Biochemical Parasitology
Título revista abreviado:Mol. Biochem. Parasitol.
ISSN:01666851
CODEN:MBIPD
CAS:etazolate, 35838-58-5, 51022-77-6; phosphatase, 9013-05-2; rolipram, 61413-54-5; sodium chloride, 7647-14-5; 3',5'-Cyclic-Nucleotide Phosphodiesterase, EC 3.1.4.17; DNA, Protozoan; Etazolate, 51022-77-6; Phosphodiesterase Inhibitors; phosphodiesterase IV, EC 3.1.4.17; Recombinant Proteins; Rolipram, 61413-54-5
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v145_n1_p40_Alonso

Referencias:

  • Mehats, C., Andersen, C.M., Filopanti, M., Jin, S.-L.C., Conti, M., Cyclic nucleotide phosphodiesterases and their role in endocrine cell signaling (2002) Trends Endocrinol Metab, 13, pp. 29-35
  • Francis, S.H., Turko, I.V., Corbin, J.D., Cyclic nucleotide phophodiesterases: Relating structure and function (2000) Prog Nucl Acids Res Mol Biol, 65, pp. 1-52
  • Ouaissi, A., Cornette, J., Schoneck, R., Fibronectin cleavage fragments provide a growth factor-like activity for the differentiation of Trypanosoma cruzi trypomastigotes to amastigotes (1992) Eur J Cell Biol, 59, pp. 68-79
  • Fraidenraich, D., Pena, C., Isola, E.L., Stimulation of Trypanosoma cruzi adenylyl cyclase by an alpha d-globin fragment from Triatoma hindgut: Effect on differentiation of epimastigote to trypomastigote forms (1993) Proc Natl Acad Sci U S a, 90, pp. 10140-10144
  • Flawiá, M.M., Tellez-Iñon, M.T., Torres, H.N., Signal transduction mechanisms in Trypanosoma cruzi (1997) Parasitol Today, pp. 1330-1333
  • Rangel-Aldao, R., Triana, F., Fernandez, V., Comach, G., Abate, T., Montoreano, R., Cyclic AMP as an inducer of the cell differentiation of Trypanosoma cruzi (1988) Biochem Int, 17, pp. 337-344
  • Naula, C., Seebeck, T., Cyclic AMP signaling in trypanosomatids (2000) Parasitol Today, 16, pp. 35-38
  • Seebeck, T., Gong, K.W., Kunz, S., Schaub, R., Shalaby, T., Zoraghi, R., Cyclic AMP signalling in Trypanosoma brucei (2001) Int J Parasitol, 31, pp. 491-498
  • Rohloff, P., Montalvetti, A., Docampo, R., Acidocalcisomes and the contractile vacuole complex are involved in osmoregulation in Trypanosoma cruzi (2004) J Biol Chem, 279, pp. 52270-52281
  • Téllez-Iñón, M.T., Ulloa, R.M., Torreula, M., Torres, H.N., Calmodulin and Ca2+-dependent cyclic AMP phosphodiesterase activity in Trypanosoma cruzi (1985) Mol Biochem Parasitol, 17, pp. 143-153
  • Walter, R.D., Opperdoes, F.R., Subcellular distribution of adenylate cyclase, cyclic-AMP phosphodiesterase, protein kinases and phosphoprotein phosphatase in Trypanosoma brucei (1982) Mol Biochem Parasitol, 6, pp. 287-295
  • Walter, R.D., 3′5′-Cyclic-AMP phosphodiesterase from Trypanosoma gambiense, Hoppe Seylers (1974) Z Physiol Chem, 355, pp. 1443-1450
  • Ráscon, A., Viloria, M.E., De-Chiara, L., Dubra, M.E., Characterization of cyclic AMP phosphodiesterases in Leishmania mexicana and purification of a soluble form (2000) Mol Biochem Parasitol, 106, pp. 283-292
  • Gong, W.K., Kunz, S., Zoraghi, R., Kunz Renggli, C., Brun, R., Seebeck, T., Cyclic AMP-specific phosphodiesterase TbPDE1 is not essential in Trypanosoma brucei in culture or during midgut infection of tsetse flies (2001) Mol Biochem Parasitol, 116, pp. 229-232
  • Zoraghi, R., Kunz, S., Gong, K., Seebeck, T., Characterization of TbPDE2A, a novel cyclic nucleotide-specific phosphodiesterase from the protozoan parasite Trypanosoma brucei (2001) J Biol Chem, 276, pp. 11559-11566
  • Zoraghi, R., Seebeck, T., The cyclic AMP-specific phosphodiesterase TbPDE2C is an essential enzyme in bloodstream form Trypanosoma brucei (2002) Proc Natl Acad Sci U S a, 99, pp. 4343-4348
  • Ráscon, A., Soderling, S.H., Schaefer, J.B., Beavo, J.A., Cloning and characterization of a cyclic AMP-specific phosphodiesterase (TbPDE2B) from Trypanosoma brucei (2002) Proc Natl Acad Sci U S a, 99, pp. 4714-4719
  • D'Angelo, M.A., Montagna, A.E., Sanguineti, S., Torres, H.N., Flawiá, M.M., A novel calcium' stimulated adenylyl cyclase from Trypanosoma cruzi which interacts with the structural flagellar protein paraflagellar rod (2002) J Biol Chem, 277, pp. 35025-35034
  • D'Angelo, M.A., Sanguineti, S., Reece, J.M., Birnbaumer, L., Torres, H.N., Flawiá, M.M., Identification, characterization and subcellular localization of TcPDE1, a novel cAMP' specific phosphodiesterase from Trypanosoma cruzi (2004) Biochem J, 378, pp. 63-72
  • Cho, W., Stahelin, R.V., Membrane-protein interactions in cell signaling and membrane trafficking (2005) Annu Rev Biophys Biomol Struct, 34, pp. 119-151
  • Blatner, N.R., Stahelin, R.V., Diraviyam, K., The molecular basis of the differential subcellular localization of FYVE domains (2004) J Biol Chem, 279, pp. 53818-53827
  • Ma, P., Wera, S., Van Dijck, P., Thevelein, J.M., The PDE1-encoded low-affinity phosphodiesterase in the yeast Saccharomyces cerevisiae has a specific function in controlling agonist-induced cyclic AMP signalling (1999) Mol Biol Cell, 10, pp. 91-104
  • Pereira, C.A., Alonso, G.D., Paveto, M.C., Trypanosoma cruzi arginine kinase characterization and cloning. a novel energetic pathway in protozoan parasites (2000) J Biol Chem, 275, pp. 1495-1501
  • Ammerer, G., Expresión of genes in yeast using the ADCI promoter (1983) Meth Enzymol, 101, pp. 192-291
  • Gietz, R.D., Schiestl, R.H., Applications of high efficiency lithium acetate transformation of intact yeast cells using single-stranded nucleic acids as carrier (1991) Yeast, 7, pp. 253-263
  • Thompson, W.J., Appleman, M.M., Multiple cyclic nucleotide phosphodiesterase activities from rat brain (1971) Biochemistry, 10, pp. 311-316
  • Londesborough, J., Quantitative estimation of 3′5′ cyclic AMP phosphodiesterase using anion exchange resin in a batch process (1976) Anal Biochem, 71, pp. 623-628
  • Laemmli, U.K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4 (1970) Nature, 227, pp. 680-685
  • Gillooly, D.J., Simonsen, A., Stenmark, H., Cellular functions of phosphatidylinositol 3-phosphate and FYVE domain proteins (2001) Biochem J, 355, pp. 249-258
  • Conti, M., Catherine Jin, S.L.C., The molecular biology of cyclic nucleotide phophodiesterases (2000) Prog Nucl Acid Res Mol Biol, 63, pp. 1-38
  • Jacobitz, S., Ryan, M.D., McLaughlin, M.M., Livi, G.P., DeWolf Jr., W.E., Torphy, T.J., Role of conserved histidines in catalytic activity and inhibitor binding in human recombinant phosphodiesterase 4A (1997) Mol Pharmacol, 51, pp. 909-1006
  • Herman, S.B., Juilfus, D.M., Fauman, E.B., Juneau, P., Menetski, J.P., Analysis of a mutation in phosphodiesterase type 4 that alters both inhibitor activity and nucleotide selectivity (2000) Mol Pharmacol, 57, pp. 991-999
  • Hayakawa, A., Hayes, S.J., Lawe, D.C., Structural basis for endosomal targeting by FYVE domains (2004) J Biol Chem, 279, pp. 5958-5966
  • Atienza, J.M., Colicelli, J., Yeast model system for study of mammalian phosphodiesterases (1998) Methods, 14, pp. 35-42
  • El-Sayed, N.M., The genome sequence of Trypanosoma cruzi, etiologic agent of Chagas Disease (2004) Science, 309, pp. 409-415
  • Torphy, T.J., Page, C., Phosphodiesterases: The journey towards therapeutics (2000) Trends Pharmacol Sci, 21, pp. 157-159

Citas:

---------- APA ----------
Alonso, G.D., Schoijet, A.C., Torres, H.N. & Flawiá, M.M. (2006) . TcPDE4, a novel membrane-associated cAMP-specific phosphodiesterase from Trypanosoma cruzi. Molecular and Biochemical Parasitology, 145(1), 40-49.
http://dx.doi.org/10.1016/j.molbiopara.2005.09.005
---------- CHICAGO ----------
Alonso, G.D., Schoijet, A.C., Torres, H.N., Flawiá, M.M. "TcPDE4, a novel membrane-associated cAMP-specific phosphodiesterase from Trypanosoma cruzi" . Molecular and Biochemical Parasitology 145, no. 1 (2006) : 40-49.
http://dx.doi.org/10.1016/j.molbiopara.2005.09.005
---------- MLA ----------
Alonso, G.D., Schoijet, A.C., Torres, H.N., Flawiá, M.M. "TcPDE4, a novel membrane-associated cAMP-specific phosphodiesterase from Trypanosoma cruzi" . Molecular and Biochemical Parasitology, vol. 145, no. 1, 2006, pp. 40-49.
http://dx.doi.org/10.1016/j.molbiopara.2005.09.005
---------- VANCOUVER ----------
Alonso, G.D., Schoijet, A.C., Torres, H.N., Flawiá, M.M. TcPDE4, a novel membrane-associated cAMP-specific phosphodiesterase from Trypanosoma cruzi. Mol. Biochem. Parasitol. 2006;145(1):40-49.
http://dx.doi.org/10.1016/j.molbiopara.2005.09.005