Subcellular localization of 14-3-3 proteins in Toxoplasma gondii tachyzoites and evidence for a lipid raft-associated form |
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Authors: | Assossou Olga Besson Françoise Rouault Jean-Pierre Persat Florence Brisson Christine Duret Laurent Ferrandiz Josette Mayençon Martine Peyron François Picot Stephane |
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Institution: | Laboratoire de Parasitologie, Mycologie Médicale et Pathologie Exotique, Equipe d'accueil (EA) 3087 Faculté de Médecine, Université Claude Bernard, 8 avenue Rockefeller, 69373, Lyon, Cedex 08, France. |
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Abstract: | A polyclonal antibody was raised against a Toxoplasma gondii 14-3-3-gluthatione S-transferase fusion protein obtained by cloning a 14-3-3 cDNA sequence determined from the T. gondii database. This antibody specifically recognized T. gondii 14-3-3 without any cross-reaction with mammalian proteins. Immunofluorescence microscopy studies of the tachyzoites or the T. gondii-infected cells suggested cytosolic and membranous localizations of 14-3-3 protein. Different subcellular fractions were prepared for electrophoresis analysis and immunodetection. 14-3-3 proteins were found in the cytosol, the membrane fraction and Triton X-100-resistant membranes. Two 14-3-3 isoforms were detected. The major one was mainly cytoplasmic and to a lesser extent membrane-associated, whereas the minor isoform was associated with the detergent-resistant lipid rafts. |
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Keywords: | 14-3-3 protein Toxoplasma gondii 14-3-3 antibody Detergent-resistant membrane Immunofluorescence microscopy Subcellular localization |
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