Molecular cloning and characterization of growth factor receptor bound-protein in Clonorchis sinensis.
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.


RESULTS
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.


CONCLUSIONS
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.
Department of Medical Environmental Biology, Chung-Ang University College of Medicine, Dongjak-gu, Seoul, Republic of Korea.
Xuelian
X
Bai
Department of Medical Environmental Biology, Chung-Ang University College of Medicine, Dongjak-gu, Seoul, Republic of Korea.
Ji-Yun
JY
Lee
Department of Medical Environmental Biology, Chung-Ang University College of Medicine, Dongjak-gu, Seoul, Republic of Korea.
Tae Im
TI
Kim
Department of Medical Environmental Biology, Chung-Ang University College of Medicine, Dongjak-gu, Seoul, Republic of Korea.
Fuhong
F
Dai
Department of Pathology, Chung-Ang Univesity College of Medicine, Dongjak-gu, Seoul, Republic of Korea.
Tae-Jin
TJ
Lee
Department of Medical Environmental Biology, Chung-Ang University College of Medicine, Dongjak-gu, Seoul, Republic of Korea.
Sung-Jong
SJ
Hong
D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D003001:Cloning, Molecular; D003003:Clonorchiasis / Q000276:immunology / Q000469:parasitology; D003004:Clonorchis sinensis / Q000235:genetics* / Q000378:metabolism; D005260:Female; D051380:GRB2 Adaptor Protein / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009928:Organ Specificity; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D011817:Rabbits; D017422:Sequence Analysis, DNA
D000947:Antigens, Helminth; D051380:GRB2 Adaptor Protein; D015801:Helminth Proteins