n- and c-terminal domains of the calcium binding protein ehcabp1 of the parasite entamoeba histolytica display distinct functions钙结合蛋白的n -和c端域ehcabp1寄生虫的痢疾阿米巴显示不同的功能.pdfVIP

N- and C-Terminal Domains of the Calcium Binding Protein EhCaBP1 of the Parasite Entamoeba histolytica Display Distinct Functions 1 1 1 2 1 Ruchi Jain , Shivesh Kumar , Samudrala Gourinath , Sudha Bhattacharya , Alok Bhattacharya * 1 School of Life Sciences, Jawaharlal Nehru University, New Delhi, India, 2 School of Environmental Sciences, Jawaharlal Nehru University, New Delhi, India Abstract Entamoeba histolytica, a protozoan parasite, is the causative agent of amoebiasis, and calcium signaling is thought to be involved in amoebic pathogenesis. EhCaBP1, a Ca2+ binding protein of E. histolytica, is essential for parasite growth. High resolution crystal structure of EhCaBP1 suggested an unusual arrangement of the EF-hand domains in the N-terminal part of the structure, while C-terminal part of the protein was not traced. The structure revealed a trimer with amino terminal domains of the three molecules interacting in a head-to-tail manner forming an assembled domain at the interface with EF1 and EF2 motifs of different molecules coming close to each other. In order to understand the specific roles of the two domains of EhCaBP1, the molecule was divided into two halves, and each half was separately expressed. The domains were characterized with respect to their structure, as well as specific functional features, such as ability to activate kinase and bind actin. The domains were also expressed in E. histolytica cells along with green fluorescent protein. The results suggest that the N-terminal domain retains some of the properties, such as localization in phagocytic cups and activation of kinase. Crystal structure of E