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Changes in cell surface proteins and glycoproteins during the encystation of Entamoeba invadens
Authors:A Chayen  B Avron  D Mirelman
Affiliation:Department of Biophysics and Unit for Molecular Biology of Parasitic Diseases, The Weizmann Institute of Science, Rehovot, Israel, 76100
Abstract:Changes in cell surface components of axenically grown trophozoites of Entamoeba invadens which occur during encystation were followed. Protein patterns of trophozoites, immature and mature cyst forms, were analyzed by sodium dodecyl sulphate gel electrophoresis. Total protein profiles of trophozoites and cyst forms stained by Coomassie blue gave similar patterns. In contrast, a number of different bands were observed in gels stained with the carbohydrate-specific Schiff's reagent as well as when nitrocellulose blottings were treated with 125I-radiolabelled wheat germ or soybean agglutinins. The most notable differences were bands at 250 and 95-105 kDa present in the cyst forms and absent in the trophozoites, and two bands at 70 and 75 kDa present in the latter and missing in the cysts. Labelling of trophozoites and cyst cell surfaces by iodination with lactoperoxidase revealed a number of protein bands which were exposed on the trophozoite surface and missing in the cysts. Moreover, gel electrophoresis patterns of non-reduced or reduced samples also differed considerably, indicating that a number of proteins are linked by disulphide bonds. This study shows that specific glycoproteins are produced during cyst formation.
Keywords:Encystation  Stage-specific proteins  Lectins  Disulphide bonds  Glycoproteins  WGA  wheat germ agglutinin  con A  concanavalin A  PAGE  polyacrylamide gel electrophoresis  PBS  phosphate buffered saline  PMSF  phenylmethylsulphonyl fluoride  SDS  sodium dodecyl sulphate  TCA  trichloroacetic acid
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