Detection and localization of actin inToxoplasma gondii
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Abstract
The immunological cross-reactivity of the protein from whole-cell extracts ofToxoplasma gondii with antisera against actins from chickengizzard smooth muscle and fromAscaris body-wall muscle was demonstrated by sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis (SDS-PAGE) and immunofluorescence studies. A protein of about 42 kDa, with the same mobility as that of rabbit smooth-muscle actin in SDS-PAGE showed cross-reactivity with anti-actin antisera on the electroblotted nitrocellulose sheet. Indirect fluorescent antibody staining of the parasites clearly showed that actin localized strongly in the anterior third of the organism in both extra- and intrahost cellular stages. From both a biochemical as well as structural point of view, the results obtained in the present experiments might explain the locomotive features of the parasite such as twirling and sliding, with most of the activity at the anterior end.
Keywords
Dodecyl Fluorescent Antibody Antibody Staining Immunofluorescence Study Structural PointPreview
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References
- Bommer W, Hofling KD, Heunert HH (1969) Multiplication ofToxoplasma gondii in cell cultures. Ger Med Monthly 14:399–405Google Scholar
- Chiappino ML, Nichols BA, O'Connor G (1984) Scanning electron microscopy ofToxoplasma gondii: parasite torsion and host cell responses during invasion. J Protozool 31:288–291PubMedGoogle Scholar
- Cintra WM, De Souza W (1985) Immunocytochemical localization of cytoskeletal proteins and electron microscopy of detergent extracted tachyzoites ofToxoplasma gondii. J Submicrosc Cytol 17:503–508PubMedGoogle Scholar
- Endo T, Tokuda H, Yagita K, Koyama T (1987) Effects of extracellular potassium on acid release and motility initiation inToxoplasma gondii. J Protozool 34:291–295PubMedGoogle Scholar
- Doran DJ (1972) Cultivation of coccidia in avian embryos and cell culture. In: Hammond DM, Long PL (eds) The coccidia. University Park Press, Baltimore, pp 183–252Google Scholar
- Jacobs L (1953) The biology ofToxoplasma. Am J Trop Med Hyg 2:365–389PubMedGoogle Scholar
- Kelley GL, Hammond DM (1967) Development ofEimeria ninakohlyakimonae from sheep in cell cultures. J Protozool 17:340–349Google Scholar
- Mühlpfordt H (1952) Das Verhalten vonToxoplasma gondii Stam BK in der Gewebekulturen. Z Tropenmed Parasitol 4:53–64PubMedGoogle Scholar
- Nakamura T, Yanagisawa T, Yamaguchi M (1975) Studies on the subunits of myosin from muscle layer ofAscaris lumbricoides suum. Biochim Biophys Acta 412:229–240PubMedGoogle Scholar
- Nakamura T, Yamaguchi M, Yanagisawa T (1979) Comparative studies on actins from various source. J Biochem 8:627–631Google Scholar
- Nichols BA, Chiappino ML (1987) Cytoskeleton ofToxoplasma gondii. J Protozool 34:217–226Google Scholar
- Nichols BA, O'Connor GR (1981) Penetration of mouse peritoneal macrophages by the protozoonToxoplasma gondii. New evidence for active invasion and phagocytosis. Lab Invest 44:324–335PubMedGoogle Scholar
- Nishioka M, Watanabe S, Kobayashi K, Nakamura T (1983) Rabbit autoantibodies to actin induced by immunization with heterologous actins; a possible mechanisms of smooth muscle antibody production. Clin Exp Immunol 5:159–164Google Scholar
- Pulvertaft RJ, Valentine JC, Lane WF (1954) The behavior ofToxoplasma gondii on serum-agar culture. Parasiology 44:478–485Google Scholar
- Russel DG, Sinden RE (1981) The role of the cytoskeleton in the motility of coccidian sporozoites. J Cell Sci 50:345–359PubMedGoogle Scholar
- Ryning FW, Remington JS (1978) Effect of cytochalasin D onToxoplasma gondii cell entry. Infect Immun 20:739–743PubMedGoogle Scholar
- Sampson JR, Hammond DM, Ernst JV (1971) Development ofEimeria alabamensis from cattle in mammalian cell cultures. J Protozool 18:120–128PubMedGoogle Scholar
- Schwartzman JD, Pfefferkorn ER (1983) Immunofluorescent localization of myosin at the anterior pole of the coccidian,Toxoplasma gondii. J Protozool 39:657–661Google Scholar
- Sheffield HG, Melton ML (1968) The fine structure and reporduction ofToxoplasma gondii. J Parasitol 54:209–226PubMedGoogle Scholar
- Sourander P, Lycke E, Lund E (1960) Observations of living cells infected withToxoplasma gondii. Br J Exp Pathol 41:176–178PubMedGoogle Scholar
- Spudich JA (1974) Biochemical and structural studies on actomyosin-like proteins from non-muscle cells. J Biol Chem 249:6013–6020PubMedGoogle Scholar
- Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354PubMedGoogle Scholar
- Trechev P, Snide P, Holborow EJ (1974) The specificity of anti-actin serum. Immunology 31:509–517Google Scholar