Subcellular localization of 14-3-3 proteins in Toxoplasma gondii tachyzoites and evidence for a lipid raft-associated form.
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.
Assossou
Olga
O
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.
Besson
Françoise
F
Rouault
Jean-Pierre
JP
Persat
Florence
F
Brisson
Christine
C
Duret
Laurent
L
Ferrandiz
Josette
J
Mayençon
Martine
M
Peyron
François
F
Picot
Stephane
S
D048948:14-3-3 Proteins; D000818:Animals; D000913:Antibodies, Protozoan; D003902:Detergents; D005455:Fluorescent Antibody Technique; D021962:Membrane Microdomains / Q000737:chemistry*; D051379:Mice; D008815:Mice, Inbred Strains; D017830:Octoxynol; D011994:Recombinant Proteins / Q000276:immunology; D014122:Toxoplasma / Q000737:chemistry* / Q000254:growth & development; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis* / Q000276:immunology
D048948:14-3-3 Proteins; D000913:Antibodies, Protozoan; D003902:Detergents; D011994:Recombinant Proteins; D017830:Octoxynol; D014446:Tyrosine 3-Monooxygenase