Antigens, Surface; Carbohydrate Biochemistry; Carbohydrate Metabolism; Cloning, Molecular; Entamoeba histolytica; Enzyme Assays; Galactose; Gene Knockdown Techniques; GPI-Linked Proteins; Kinetics; Microbiology; Molecular Biology; Parasitology; Peptidoglycan; UDPglucose 4-Epimerase
The intestinal protozoan parasite Entamoeba histolytica still claims more than 50,000 human lives every year. Our work aims to understand the antigenic structures of the parasite on the one hand to view them as vaccine candidates and on the other hand to understand their biosynthesis to identify new ways to interfere with the parasite. In a current project we collaborate with the lab of Iain Wilson at the BOKU Vienna. In this project funded by the FWF we returned to a very complex surface antigen, called lipopeptidophosphoglycan (LPPG), against which our lab had once raised a monoclonal antibody. One aim is to identify the suspected core protein component, the other aim is to unravel the bioythesis of LPPG and its precursors. Galactose is an important sugar in the glycan part of LPPG, and E. histolytica displays intesting pathways to make use of this sugar.
Techniques, methods & infrastructure
The parasite Entamoeba histolytica is cultured routinely in our lab. One aim of the current project is to examine whether the product of the identified gene is really contained in the LPPG structure. The other part of the project aims at the biosynthesis of the structure and its precursors. This is done by conventional molecular cloning and expression methods as well as two-dimensional electrophoreses and identification of spots by mass spectrometry. Enzyme products are examined by HPLC in the lab of our collaborator Iain Wilson. In addition, specific knockdown of selected genes in E. histolytica is attempted.