Subcellular fractionation and molecular characterization of the pellicle and plasmalemma of Neospora caninum
- 16 June 2005
- journal article
- Published by Cambridge University Press (CUP) in Parasitology
- Vol. 131 (04), 467-475
- https://doi.org/10.1017/s003118200500805x
Abstract
A characteristic structural feature of Toxoplasma gondii and Neospora caninum is the presence of a triple-membrane pellicle, on the zoite stages of their complex life-cycle. Here we report the results of electron microscopic studies which show that the pellicle is made of a typical plasmalemma covered on its cytoplasmic side by a system of flattened vesicles named the inner membrane complex. Using methods described previously for the purification of pellicle and plasmalemma fractions from T. gondii, we have evaluated the same methodology for the preparation of pellicles and plasmalemma from N. caninum. The approach used involved subcellular fractionation and sucrose gradient centrifugation to prepare fractions containing pellicles. Plasmalemma was prepared by extraction of this fraction with a high salt glycerol treatment. Fractions containing membrane structures were identified by electron microscopy, and the proteins and antigens present in them were subsequently studied by SDS-PAGE and Western blotting. Electron microscopy of the pellicle fractions of N. caninum demonstrated preservation of the triple-membrane structure which is identical to that found in T. gondii. SDS-PAGE of the pellicle fractions revealed it contained several major proteins. Analyses revealed that the plasmalemma of N. caninum contained 2 abundant proteins in addition to other much lower abundance antigens detectable by monoclonal antibodies. These studies therefore report, for the first time, a detailed molecular characterization of the pellicle and plasmalemma of N. caninum.Keywords
This publication has 30 references indexed in Scilit:
- The SRS superfamily of Toxoplasma surface proteinsInternational Journal for Parasitology, 2004
- Neospora caninum: Identification of 19-, 38-, and 40-kDa Surface Antigens and a 33-kDa Dense Granule Antigen Using Monoclonal AntibodiesExperimental Parasitology, 1999
- Identification of a major surface protein on Neospora caninum tachyzoitesZeitschrift Fur Parasitenkunde-Parasitology Research, 1996
- Isolation of the pellicle ofToxoplasma gondii (Protozoa, Coccidia): Characterization by electron microscopy and protein compositionZeitschrift Fur Parasitenkunde-Parasitology Research, 1990
- Surface antigens ofToxoplasma gondiiParasitology, 1988
- Detection and localization of actin inToxoplasma gondiiZeitschrift Fur Parasitenkunde-Parasitology Research, 1988
- Cytoskeleton of Toxoplasma gondii1The Journal of Protozoology, 1987
- Monoclonal Antibodies to Toxoplasma Cell Membrane Surface Antigens Protect Mice from Toxoplasmosis1,2The Journal of Protozoology, 1983
- Toxoplasma gondii: Ultrastructure and antigenicity of purified tachyzoite pellicleExperimental Parasitology, 1979
- The Fine Structure and Mode of Division of Toxoplasma gondiiArchives of Ophthalmology (1950), 1966