Identification, characterization and subcellular localization of TcPDE1, a novel cAMP-specific phosphodiesterase from Trypanosoma cruzi

被引:29
作者
D'Angelo, MA
Sanguineti, S
Reece, JM
Birnbaumer, L
Torres, HN
Flawiá, MM
机构
[1] Univ Buenos Aires, Inst Invest & Ingn Genet & Biol Mol, Consejo Nacl Invest Cient & Tecn, RA-1428 Buenos Aires, DF, Argentina
[2] Univ Buenos Aires, Fac Ciencias Exactas & Nat, RA-1428 Buenos Aires, DF, Argentina
[3] NIEHS, Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA
关键词
cAMP; flagellum; membrane; phosphodiesterase; Trypanosoma cruzi;
D O I
10.1042/BJ20031147
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Compartmentalization of cAMP phosphodiesterases plays a key role in the regulation of cAMP signalling in mammals. In the present paper, we report the characterization and subcellular localization of TcPDE1, the first cAMP-specific phosphodiesterase to be identified from Trypanosoma cruzi. TcPDE1 is part of a small gene family and encodes a 929-amino-acid protein that can complement a heat-shock-sensitive yeast mutant deficient in phosphodiesterase genes. Recombinant TcPDE1 strongly associates with membranes and cannot be released with NaCl or sodium cholate, suggesting that it is an integral membrane protein. This enzyme is specific for cAMP and its activity is not affected by cGMP. Ca2+, calmodulin or fenotiazinic inhibitors. TcPDE1 is sensitive to the phosphodiesterase inhibitor dipyridamole but is resistant to 3-isobutyl-1-methylxanthine, theophylline, rolipram and zaprinast. Papaverine, erythro-9-(2-hydroxy-3-nonyl)-adenine hydrochloride, and vinpocetine are poor inhibitors of this enzyme. Confocal laser scanning of T. cruzi epimastigotes showed that TcPDE1 is associated with the plasma membrane and concentrated in the flagellum of the parasite. The association of TcPDE1 with this organelle was confirmed by subcellular fractionation and cell-disruption treatments. The localization of this enzyme is a unique feature that distinguishes it from all the trypanosomatid phosphodiesterases described so far and indicates that compartmentalization of cAMP phosphodiesterases could also be important in these parasites.
引用
收藏
页码:63 / 72
页数:10
相关论文
共 47 条
[1]   Yeast model system for study of mammalian phosphodiesterases [J].
Atienza, JM ;
Colicelli, J .
METHODS, 1998, 14 (01) :35-42
[2]   In addition to the SH3 binding region, multiple regions within the N-terminal noncatalytic portion of the cAMP-specific phosphodiesterase, PDE4A5, contribute to its intracellular targeting [J].
Beard, MB ;
Huston, E ;
Campbell, L ;
Gall, I ;
McPhee, I ;
Yarwood, S ;
Scotland, G ;
Houslay, MD .
CELLULAR SIGNALLING, 2002, 14 (05) :453-465
[3]   MODIFIED ASSAY OF 3'-5'-CYCLIC-AMP PHOSPHODIESTERASE [J].
BOUDREAU, RJ ;
DRUMMOND, GI .
ANALYTICAL BIOCHEMISTRY, 1975, 63 (02) :388-399
[4]   ISOLATION AND CHARACTERIZATION OF A MAMMALIAN GENE ENCODING A HIGH-AFFINITY CAMP PHOSPHODIESTERASE [J].
COLICELLI, J ;
BIRCHMEIER, C ;
MICHAELI, T ;
ONEILL, K ;
RIGGS, M ;
WIGLER, M .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (10) :3599-3603
[5]   AKAPs: from structure to function [J].
Colledge, M ;
Scott, JD .
TRENDS IN CELL BIOLOGY, 1999, 9 (06) :216-221
[6]   A novel calcium-stimulated adenylyl cyclase from Trypanosoma cruzi, which interacts with the structural flagellar protein paraflagellar rod [J].
D'Angelo, M ;
Montagna, AE ;
Sanguineti, S ;
Torres, HN ;
Flawiá, MM .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (38) :35025-35034
[7]  
Ersfeld K, 2001, J CELL SCI, V114, P141
[8]   Signal transduction mechanisms in Trypanosoma cruzi [J].
Flawia, MM ;
TellezInon, MT ;
Torres, HN .
PARASITOLOGY TODAY, 1997, 13 (01) :30-33
[9]   Solubilization of membrane-bound rod phosphodiesterase by the rod phosphodiesterase recombinant delta subunit [J].
Florio, SK ;
Prusti, RK ;
Beavo, JA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (39) :24036-24047
[10]   STIMULATION OF TRYPANOSOMA-CRUZI ADENYLYL-CYCLASE BY AN ALPHA(D)-GLOBIN FRAGMENT FROM TRIATOMA-HINDGUT - EFFECT ON DIFFERENTIATION OF EPIMASTIGOTE TO TRYPOMASTIGOTE FORMS [J].
FRAIDENRAICH, D ;
PENA, C ;
ISOLA, EL ;
LAMMEL, EM ;
COSO, O ;
ANEL, AD ;
PONGOR, S ;
BARALLE, F ;
TORRES, HN ;
FLAWIA, MM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (21) :10140-10144