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Molecular cloning and characterization of growth factor receptor bound-protein in Clonorchis sinensis.
- Source :
-
PloS one [PLoS One] 2014 Jan 16; Vol. 9 (1), pp. e85577. Date of Electronic Publication: 2014 Jan 16 (Print Publication: 2014). - Publication Year :
- 2014
-
Abstract
- Background: Clonorchis sinensis causes clonorchiasis, a potentially serious disease. Growth factor receptor-bound protein 2 (Grb2) is a cytosolic protein conserved among animals and plays roles in cellular functions such as meiosis, organogenesis and energy metabolism. In the present study, we report first molecular characters of growth factor receptor bound-protein (CsGrb2) from C. sinensis as counter part of Grb2 from animals and its possible functions in development and organogenesis of C. sinensis.<br />Methodology/principal Findings: A CsGrb2 cDNA clone retrieved from the C. sinensis transcriptome encoded a polypeptide with a SH3-SH2-SH3 structure. Recombinant CsGrb2 was bacterially produced and purified to homogeneity. Native CsGrb2 with estimated molecular weight was identified from C. sinensis adult extract by western blotting using a mouse immune serum to recombinant CsGrb2. CsGrb2 transcripts was more abundant in the metacercariae than in the adults. Immunohistochemical staining showed that CsGrb2 was localized to the suckers, mesenchymal tissues, sperms in seminal receptacle and ovary in the adults, and abundantly expressed in most organs of the metacercariae. Recombinant CsGrb2 was evaluated to be little useful as a serodiagnostic reagent for C. sinesis human infections.<br />Conclusion: Grb2 protein found in C. sinensis was conserved among animals and suggested to play a role in the organogenesis, energy metabolism and mitotic spermatogenesis of C. sinensis. These findings from C. sinensis provide wider understanding on diverse function of Grb2 in lower animals such as platyhelminths.
- Subjects :
- Amino Acid Sequence
Animals
Antigens, Helminth chemistry
Antigens, Helminth genetics
Antigens, Helminth immunology
Antigens, Helminth metabolism
Cloning, Molecular
Clonorchiasis immunology
Clonorchiasis parasitology
Clonorchis sinensis metabolism
Female
GRB2 Adaptor Protein chemistry
GRB2 Adaptor Protein immunology
GRB2 Adaptor Protein metabolism
Gene Expression Regulation, Developmental
Helminth Proteins chemistry
Helminth Proteins immunology
Helminth Proteins metabolism
Humans
Male
Mice
Mice, Inbred BALB C
Molecular Sequence Data
Molecular Weight
Organ Specificity
Phylogeny
Protein Structure, Tertiary
Rabbits
Sequence Analysis, DNA
Clonorchis sinensis genetics
GRB2 Adaptor Protein genetics
Helminth Proteins genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 9
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 24454892
- Full Text :
- https://doi.org/10.1371/journal.pone.0085577