The Journal of Immunology, Vol 130, Issue 5 2407-2412, Copyright © 1983 by American Association of Immunologists

ARTICLES

Purification of a major membrane protein of Toxoplasma gondii by immunoabsorption with a monoclonal antibody

LH Kasper, JH Crabb and ER Pfefferkorn

The principal iodinatable surface protein (P30) of our cloned RH strain of Toxoplasma gondii has an apparent molecular weight of 30,000, as measured by acrylamide gel electrophoresis in the presence of sodium dodecyl sulfate under reducing conditions. Monoclonal antibody B specifically immunoprecipitated protein P30 from a detergent extract of surface radioiodinated T. gondii. Monoclonal antibody B in the presence of complement was also parasiticidal for T. gondii, and this parasiticidal effect could be blocked by protein P30. Monoclonal antibody B was purified from mouse ascitic fluid and linked to cyanogen bromide-activated Sepharose. The resulting immunoabsorbent was used to purify 1.7 mg of protein P30 from a large number of parasites. The efficiency of recovery of protein P30 was measured by assays of radioactivity and of parasiticidal blocking activity. Protein P30 represented 3 to 5% of the total protein. It is also present in a recently isolated strain of T. gondii. A convalescent human antitoxoplasma serum immunoprecipitated radiolabeled protein P30. Three convalescent antisera when quantitated by an ELISA test had a high anti- protein P30 titer. Charge shift electrophoresis showed that protein P30 has an extensive hydrophobic region and thus is probably an integral membrane protein. Electrophoresis under nonreducing conditions showed no evidence that protein P30 exists as a disulfide linked homo- or heterodimer, although it probably has intramolecular disulfide bonds.

This article has been cited by other articles:

				P. J. Bradley, C. Ward, S. J. Cheng, D. L. Alexander, S. Coller, G. H. Coombs, J. D. Dunn, D. J. Ferguson, S. J. Sanderson, J. M. Wastling, et al. Proteomic Analysis of Rhoptry Organelles Reveals Many Novel Constituents for Host-Parasite Interactions in Toxoplasma gondii J. Biol. Chem., October 7, 2005; 280(40): 34245 - 34258. [Abstract] [Full Text] [PDF]

				N. Rachinel, D. Buzoni-Gatel, C. Dutta, F. J. D. Mennechet, S. Luangsay, L. A. Minns, M. E. Grigg, S. Tomavo, J. C. Boothroyd, and L. H. Kasper The Induction of Acute Ileitis by a Single Microbial Antigen of Toxoplasma gondii J. Immunol., August 15, 2004; 173(4): 2725 - 2735. [Abstract] [Full Text] [PDF]

				V. Letscher-Bru, A. W. Pfaff, A. Abou-Bacar, D. Filisetti, E. Antoni, O. Villard, J.-P. Klein, and E. Candolfi Vaccination with Toxoplasma gondii SAG-1 Protein Is Protective against Congenital Toxoplasmosis in BALB/c Mice but Not in CBA/J Mice Infect. Immun., November 1, 2003; 71(11): 6615 - 6619. [Abstract] [Full Text] [PDF]

				M. J. Wichroski, J. A. Melton, C. G. Donahue, R. K. Tweten, and G. E. Ward Clostridium septicum Alpha-Toxin Is Active against the Parasitic Protozoan Toxoplasma gondii and Targets Members of the SAG Family of Glycosylphosphatidylinositol-Anchored Surface Proteins Infect. Immun., August 1, 2002; 70(8): 4353 - 4361. [Abstract] [Full Text] [PDF]

				M. Marti, Y. Li, P. Kohler, and A. B. Hehl Conformationally Correct Expression of Membrane-Anchored Toxoplasma gondii SAG1 in the Primitive Protozoan Giardia duodenalis Infect. Immun., February 1, 2002; 70(2): 1014 - 1016. [Abstract] [Full Text] [PDF]

				C. Dutta, J. Grimwood, and L. H. Kasper Attachment of Toxoplasma gondii to a Specific Membrane Fraction of CHO Cells Infect. Immun., December 1, 2000; 68(12): 7198 - 7201. [Abstract] [Full Text] [PDF]

				M. W. Black and J. C. Boothroyd Lytic Cycle of Toxoplasma gondii Microbiol. Mol. Biol. Rev., September 1, 2000; 64(3): 607 - 623. [Abstract] [Full Text] [PDF]

				C. D. Jones, N. Okhravi, P. Adamson, S. Tasker, and S. Lightman Comparison of PCR Detection Methods for B1, P30, and 18S rDNA Genes of T. Gondii in Aqueous Humor Invest. Ophthalmol. Vis. Sci., March 1, 2000; 41(3): 634 - 644. [Abstract] [Full Text]

				F. Velge-Roussel, P. Marcelo, A. C. Lepage, D. Buzoni-Gatel, and D. T. Bout Intranasal Immunization with Toxoplasma gondii SAG1 Induces Protective Cells into Both NALT and GALT Compartments Infect. Immun., February 1, 2000; 68(2): 969 - 972. [Abstract] [Full Text] [PDF]

				J. Y. Channon, E. I. Suh, R. M. Seguin, and L. H. Kasper Attachment Ligands of Viable Toxoplasma gondii Induce Soluble Immunosuppressive Factors in Human Monocytes Infect. Immun., May 1, 1999; 67(5): 2547 - 2551. [Abstract] [Full Text] [PDF]

				V. Letscher-Bru, O. Villard, B. Risse, M. Zauke, J.-P. Klein, and T. T. Kien Protective Effect of Vaccination with a Combination of Recombinant Surface Antigen 1 and Interleukin-12 against Toxoplasmosis in Mice Infect. Immun., September 1, 1998; 66(9): 4503 - 4506. [Abstract] [Full Text] [PDF]

				N. M. Silva, R. T. Gazzinelli, D. A. O. Silva, E. A. V. Ferro, L. H. Kasper, and J. R. Mineo Expression of Toxoplasma gondii-Specific Heat Shock Protein 70 during In Vivo Conversion of Bradyzoites to Tachyzoites Infect. Immun., August 1, 1998; 66(8): 3959 - 3963. [Abstract] [Full Text] [PDF]

				J. Hackett Jr., J. Hoff-Velk, A. Golden, J. Brashear, J. Robinson, M. Rapp, M. Klass, D. H. Ostrow, and W. Mandecki Recombinant Mouse-Human Chimeric Antibodies as Calibrators in Immunoassays That Measure Antibodies to Toxoplasma gondii J. Clin. Microbiol., May 1, 1998; 36(5): 1277 - 1284. [Abstract] [Full Text]

				L. J. Knoll and J. C. Boothroyd Isolation of Developmentally Regulated Genes from Toxoplasma gondii by a Gene Trap with the Positive and Negative Selectable Marker Hypoxanthine-Xanthine-Guanine Phosphoribosyltransferase Mol. Cell. Biol., February 1, 1998; 18(2): 807 - 814. [Abstract] [Full Text]

				K. Joiner, S. Fuhrman, H. Miettinen, L. Kasper, and I Mellman Toxoplasma gondii: fusion competence of parasitophorous vacuoles in Fc receptor-transfected fibroblasts Science, August 10, 1990; 249(4969): 641 - 646. [Abstract] [PDF]