Identification of novel cyclic nucleotide binding proteins in Trypanosoma cruzi.
Cyclic AMP has been implicated as second messenger in a wide range of cellular processes. In the protozoan parasite Trypanosoma cruzi, cAMP is involved in the development of the parasite's life cycle. While cAMP effectors have been widely studied in other eukaryotic cells, little is known about cAMP's mechanism of action in T. cruzi. To date, only a cAMP-dependent protein kinase A (PKA) has been cloned and characterised in this parasite; however experimental evidence indicates the existence of cAMP-dependent, PKA-independent events. In order to identify new cAMP binding proteins as potential cAMP effectors, we carried out in silico studies using the predicted T. cruzi proteome. Using a combination of search methods 27 proteins with putative cNMP binding domains (CBDs) were identified. Phylogenetic analysis of the CBDs presented a homogeneous distribution, with sequences segregated into two main branches: one containing kinases-like proteins and the other gathering hypothetical proteins with different function or no other known. Comparative modelling of the strongest candidates provides support for the hypothesis that these proteins may give rise to structurally viable cyclic nucleotide binding domains. Pull-down and nucleotide displacement assays strongly suggest that TcCLB.508523.80 could bind cAMP and eventually be a new putative PKA-independent cAMP effector in T. cruzi.
Instituto de Investigaciones Biotecnológicas-Instituto Tecnológico de Chascomús, UNSAM-CONICET, Buenos Aires, Argentina.
Adriana V
AV
Jäger
Instituto de Investigaciones Biotecnológicas-Instituto Tecnológico de Chascomús, UNSAM-CONICET, Buenos Aires, Argentina.
Javier G
JG
De Gaudenzi
Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad de Buenos Aires, Argentina; IQUIBICEN-CONICET, Ciudad de Buenos Aires, Argentina.
Jesica G
JG
Mild
Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad de Buenos Aires, Argentina.
Bárbara
B
Mc Cormack
Institut Pasteur de Montevideo, Uruguay.
Sergio
S
Pantano
Department of Pharmacology and Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA. Electronic address: altschul@pitt.edu.
Daniel L
DL
Altschuler
Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad de Buenos Aires, Argentina; IQUIBICEN-CONICET, Ciudad de Buenos Aires, Argentina; Department of Pharmacology and Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA. Electronic address: mme2@pitt.edu.
Martin M
MM
Edreira
D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D016000:Cluster Analysis; D019295:Computational Biology; D009712:Nucleotides, Cyclic / Q000378:metabolism*; D010802:Phylogeny; D011485:Protein Binding; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D014349:Trypanosoma cruzi / Q000235:genetics / Q000378:metabolism*
D002352:Carrier Proteins; D009712:Nucleotides, Cyclic; D015800:Protozoan Proteins
Trypanosoma cruzi; cAMP binding proteins; cAMP novel effectors; cAMP signalling