Kim Tae Im, Cho Pyo Yun, Li Shunyu, Hong Sung-Tae, Choi Min-Ho, Hong Sung-Jong
Department of Parasitology, Chung-Ang University College of Medicine, Tongjak-gu, Seoul, 156-756, South Korea.
Parasitol Res. 2007 Oct;101(5):1233-8. doi: 10.1007/s00436-007-0625-5. Epub 2007 Jul 6.
WD40-repeat proteins have four to eight repeat units, which have Gly-His (GH) and Trp-Asp (WD) at both termini and fold into a beta-propeller. In particular, the WD40-repeat protein of Clonorchis sinensis (CsWD1) has seven WD-repeat units and is expressed stage-specifically in metacercariae. By yeast two-hybrid screening, putative interacting protein cDNAs were cloned from a C. sinensis metacercaria cDNA library and purified further by higher stringency screening and lacZ colony-lift assay. After assessing their nucleotide and polypeptide sequences, 21 putative partner protein cDNAs were selected and assembled into 14 clones. Using YRG2 strain yeast, 12 putative partner protein clones were confirmed to interact with CsWD1 protein. These 12 proteins were grouped into functional categories, i.e., signal proteins, transporters, proteases, and muscle proteins. These results suggest that CsWD1 protein is associated with intracellular protein translocation and cell cycle control in C. sinensis metacercaria.
WD40重复蛋白具有4至8个重复单元,其两端均有甘氨酸-组氨酸(GH)和色氨酸-天冬氨酸(WD),并折叠成β-螺旋桨结构。特别地,华支睾吸虫的WD40重复蛋白(CsWD1)有7个WD重复单元,且在尾蚴中呈阶段特异性表达。通过酵母双杂交筛选,从华支睾吸虫尾蚴cDNA文库中克隆出推定的相互作用蛋白cDNA,并通过更高严谨度筛选和lacZ菌落转移试验进一步纯化。在评估其核苷酸和多肽序列后,选择了21个推定的伴侣蛋白cDNA并组装成14个克隆。使用YRG2菌株酵母,证实12个推定的伴侣蛋白克隆与CsWD1蛋白相互作用。这12种蛋白被归类为功能类别,即信号蛋白、转运蛋白、蛋白酶和肌肉蛋白。这些结果表明,CsWD1蛋白与华支睾吸虫尾蚴的细胞内蛋白转运和细胞周期调控有关。