Back to Search
Start Over
Cloning and in vitro expression of TPK3, a Toxoplasma gondii homologue of shaggy/glycogen synthase kinase-3 kinases.
- Source :
-
Molecular and biochemical parasitology [Mol Biochem Parasitol] 1998 Jun 01; Vol. 93 (2), pp. 273-83. - Publication Year :
- 1998
-
Abstract
- As an initial effort to dissect the signaling pathways responsible for pathogenesis of Toxoplasma gondii infection, we report the cloning and in vitro functional studies of TPK3 (Toxoplasma protein kinase-3), a homologue of shaggy/glycogen synthase kinase-3 (GSK-3) kinases. The shaggy/GSK-3 family of kinases are highly conserved protein kinases that play important roles in cell fate determination, nuclear signaling and hormonal regulation. The TPK3 gene was isolated by RT-PCR with degenerate primers corresponding to highly conserved regions of serine/threonine protein kinases. The complete sequences of genomic and cDNA clones indicated the open reading frame, 1185 bp in size, is interrupted by five introns. The predicted protein sequence of TPK3 shows 54% identity to shaggy/GSK-3 over the catalytic domains. Southern analysis revealed TPK3 is a single copy locus in the Toxoplasma genome. Antisera to other GSK-3 proteins from other species recognized GST-TPK3 and a protein of the predicted size in parasites lysates. In vitro kinase assays with GST-TPK3 indicated that TPK3 autophosphorylates and phosphorylates protein phosphatase inhibitor-2 (I-2), a specific substrate of GSK-3 kinase.
- Subjects :
- Amino Acid Sequence
Animals
Base Sequence
Blotting, Northern
Blotting, Southern
Blotting, Western
Calcium-Calmodulin-Dependent Protein Kinases chemistry
Calcium-Calmodulin-Dependent Protein Kinases metabolism
Cloning, Molecular
DNA, Complementary
Gene Expression
Glycogen Synthase Kinase 3
Humans
Molecular Sequence Data
Phosphorylation
Protein Serine-Threonine Kinases chemistry
Protein Serine-Threonine Kinases genetics
Recombinant Fusion Proteins metabolism
Sequence Alignment
Sequence Analysis, DNA
Substrate Specificity
Toxoplasma enzymology
Calcium-Calmodulin-Dependent Protein Kinases genetics
Drosophila Proteins
Genes, Protozoan
Protozoan Proteins
Toxoplasma genetics
Subjects
Details
- Language :
- English
- ISSN :
- 0166-6851
- Volume :
- 93
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Molecular and biochemical parasitology
- Publication Type :
- Academic Journal
- Accession number :
- 9662711
- Full Text :
- https://doi.org/10.1016/s0166-6851(98)00042-5