Abstract
Surface proteins ofSarcocystis cyst merozoites were labeled by biotinylation or radioiodination and identified on Western blots after sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). The major labeled proteins ofS. muris andS. suicanis have relative molecular masses of 31 and 33 kDa, respectively. Immunoblots performed with the 31-kDa protein and sera of experimentally infected mice or with monoclonal antibodies toS. muris revealed that this protein is immunogenic. Indirect fluorescent antibody tests (IFAT) executed usingSarcocystis cyst merozoites and polyclonal monospecific antibodies obtained from rabbits immunized with homogeneous major surface antigens gave additional evidence for the localization of the identified antigens in the pellicle. Analysis ofS. muris andS. suicanis proteins by two-dimensional gel electrophoresis revealed multiple isoelectric forms.
Similar content being viewed by others
References
Abbas MK, Powell EC (1983) Identification of surface antigens ofSarcocystis muris (Coccidia). J Protozool 30:356–361
Augustine PC (1989) Molecular interactions of cultured turkey kidney cells with specific antigens ofEimeria adenoeides sporozoites. PSEBM 191:30–36
Augustine PC, Danforth HD (1985) Effects of hybridoma antibodies on invasion of cultured cells by sporozoites ofEimeria. Avian Dis 29:1212–1223
Burgess DE, Speer CA, Reduker DW (1988) Identification of antigens ofSarcocystis cruzi sporozoites merozoites, and bradyzoites with monoclonal antibodies. J Parasitol 74:828–832
Chbouki N, Dubremetz JF (1985) Structure isolation and protein composition of the pellicle ofSarcocystis muris cystozoites (Protozoa, Coccidia). J Protozool 32:54–58
Cole SR, Ashman LK, Ey PL (1987) Biotinylation: an alternative to radioiodination for the identification of cell surface antigens in immunoprecipitates. Mol Immunol 24:699–705
Dubremetz JF (1989) Motility and penetration. In: Mehlhorn H (ed) Parasitology in focus. Springer: Berlin Heidelberg New York, pp 188–196
Fraker PM, Speck JC (1978) Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3α, 6α-diphenylglycoluril. Biochem Biophys Res Commun 80:849–857
Hancock K, Tsang VCW (1983) India ink staining of proteins on nitrocellulose paper. Anal Biochem 133:157–162
Howard RJ, Kaushal DC, Carter R (1982) Radioiodination of parasite antigens with 1,3,4,6-tetracloro-3α, 6α-diphenylglycoluril (Iodogen): studies with zygotes ofPlasmodium gallinaceum. J Protozool 29:114–117
Hurley WL, Finkelstein E, Holst BD (1985) Identification of surface proteins on bovine leucocytes by a biotin-avidin blotting technique. J Immunol Methods 85:195–202
Kasper LH, Crabb JH, Pfefferkorn ER (1983) Purification of a major membrane protein ofToxoplasma gondii by immunoabsorption with monoclonal antibody. J Immunol 130:2407–2412
Kyhse-Andersen J (1984) Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Methods 10:203–209
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lai CY, Chen C, Horecker BL (1970) Primary structure of two COOH-terminal hexapeptides from rabbit muscle aldolase: a difference in the structure of the α and β subunits. Biochem Biophys Res Commun 40:461–468
Laing P, Parkar BA, Culbert EJ, Watt GJ, Elson CJ (1987) Identification of rat erythrocyte antigens with a new non-radioactive immunoprecipitation technique. Scand J Immunol 25:613–620
Lima MF, Villalta F (1988) Host-cell attachment byTrypanosoma cruzi: identification of an adhesion molecule. Biochem Biophys Res Commun 155:256–262
Lima MF, Villalta F (1989)Trypanosoma cruzi trypomastigote clones differentially express a parasite cell adhesion molecule. Mol Biochem Parasitol 33:159–170
Lunde MN, Fayer R (1977) Serologic tests for antibody toSarcocystis in cattle. J Parasitol 63:222–225
Markwell MAK (1982) A new solid-state reagent to iodinate proteins. Anal Biochem 125:427–432
Mehlhorn H, Heydorn AO (1978) The Sarcosporidia: life cycle and fine structure. Adv Parasitol 16:43–103
Mencke N (1989) Untersuchung von Sarkosporidien-Zystozoiten in der Immuno-Elektrophorese (Western Blot) und der Immuno-Ekeltronenmikroskopie mit Hilfe von monoklonnalen Antikörpern gegenSarcocystis muris. Thesis, Tierärztliche Hochschule, Hannover
Morrissey JH (1981) Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem 117:307–310
O'Donoghue PJ, Weyreter H (1984) Examinations on the serodiagnosis ofSarcocystis infections: II. Class-specific immunoglobulin responses in mice, pigs and sheep. Zentralbl Bakteriol Mikrobiol Hyg [A] 257:168–184
O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021
Richardson K, Parker CD (1985) Identification and characterization ofVibrio cholerae surface proteins by radioiodination. Infect Immun 48:87–93
Ruiz A, Frenkel JK (1976) Recognition of cyclic transmissionSarcocystis muris by cats. J Infect Dis 133:527–531
Salacinski PRP, McLean C, Sykes JEC, Clement-Jones VV, Lowry PJ (1981) Iodination of proteins, glycoproteins, and peptides using a solid-phase oxidizing agent, 1,3,4,6-tetrachloro-3α,6α-diphenyl glycoluril (Iodogen). Anal Biochem 117:136–146
Tanabe K, Takasaki S, Watanabe JI, Kobata A, Egawa K Nakamura Y (1989) Glycoproteins composed of major surface immunodeterminants ofPneumocystis carinii. Infect Immuno 57:1363–1368
Tenter AM, Vietmeyer C, Thümmel P, Rommel M (1991) Characterization of hybridoma antibodies toSarcocystis muris. Parasitol Res (in press)
Author information
Authors and Affiliations
Additional information
Supported by the Deutsche Forschungsgemeinschaft and dedicated to Prof. Dr. Johannes Eckert (Zürich) on the occasion of his 60th birthday
Rights and permissions
About this article
Cite this article
Sommer, I., Mehlhorn, H. & Rüger, W. Biochemical and immunological characterization of major surface antigens ofSarcocystis muris andS. suicanis cyst merozoites. Parasitol Res 77, 204–211 (1991). https://doi.org/10.1007/BF00930859
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00930859