Abstract
A ribosomal fraction of Trypanosoma cruzi was isolated with detergent lysis and differential ultracentrifugation. This ribosomal fraction directed in vitro protein synthesis, in a heterologous incorporation system (rat liver pH 5 fraction), leading to values up to 10 times higher than endogenous control. The ideal concentrations of Mg2+, K+, and ribosomal proteins and the time of incubation of the incorporation mixtures were 6 mM, 21 mM, 60 microliters, and 45 min, respectively. The product thus obtained was analyzed by fluorography after polyacrylamide-sodium dodecyl sulfate gel electrophoresis and showed the presence of many protein bands, but few bands were present in the immunoprecipitate obtained with serum from Chagas' disease patients. This product was able to react with anti-T. cruzi antibodies when submitted to an immunoenzymatic assay.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Avrameas S. Coupling of enzymes to proteins with glutaraldehyde. Use of the conjugates for the detection of antigens and antibodies. Immunochemistry. 1969 Jan;6(1):43–52. doi: 10.1016/0019-2791(69)90177-3. [DOI] [PubMed] [Google Scholar]
- Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
- Castro C., Hernández R., Castañeda M. Trypanosoma cruzi ribosomal RNA: internal break in the large-molecular-mass species and number of genes. Mol Biochem Parasitol. 1981 Feb;2(3-4):219–233. doi: 10.1016/0166-6851(81)90102-x. [DOI] [PubMed] [Google Scholar]
- Crane M. S., Dvorak J. A. Trypanosoma cruzi: pattern of RNA synthesis following infection of vertebrate cells. J Protozool. 1980 Aug;27(3):336–338. doi: 10.1111/j.1550-7408.1980.tb04273.x. [DOI] [PubMed] [Google Scholar]
- Eggitt M. J., Tappenden L., Brown K. N. Translation in a reticulocyte cell-free system of RNA isolated from blood and culture forms of Trypanosoma brucei. Parasitology. 1977 Oct;75(2):133–141. doi: 10.1017/s0031182000062272. [DOI] [PubMed] [Google Scholar]
- Goldberg I. H., Mitsugi K. Sparsomycin inhibition of polypeptide synthesis promoted by synthetic and natural polynucleotides. Biochemistry. 1967 Feb;6(2):372–382. doi: 10.1021/bi00854a002. [DOI] [PubMed] [Google Scholar]
- Huang M. T., Grollman A. P. Pyrocatechol violet: an inhibitor of initiation of protein synthesis. Biochem Biophys Res Commun. 1973 Aug 21;53(4):1049–1059. doi: 10.1016/0006-291x(73)90571-8. [DOI] [PubMed] [Google Scholar]
- Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
- Lheureux M., Lheureux M., Vervoort T., Van Meirvenne N., Steinert M. Immunological purification and partial characterization of variant-specific surface antigen messenger RNA of Trypanosoma brucei brucei. Nucleic Acids Res. 1979 Oct 10;7(3):595–609. doi: 10.1093/nar/7.3.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Merritt S. C. Purification and cell-free translation of mRNA coding for a variant specific antigen from Trypanosoma brucei gambiense. Mol Biochem Parasitol. 1980 Jun;1(3):151–166. doi: 10.1016/0166-6851(80)90014-6. [DOI] [PubMed] [Google Scholar]
- Morales N. M., Roberts J. F. The ribonucleic acids of Crithidia fasciculata. J Protozool. 1978 Feb;25(1):140–144. doi: 10.1111/j.1550-7408.1978.tb03886.x. [DOI] [PubMed] [Google Scholar]
- Ono Y., Skoultchi A., Waterson J., Lengyel P. Peptide chain elongation: GTP cleavage catalysed by factors binding aminoacyl-transfer RNA to the ribosome. Nature. 1969 May 17;222(5194):645–648. doi: 10.1038/222645a0. [DOI] [PubMed] [Google Scholar]
- Rondinelli E., Soares M. C., de Castro J. F., de Castro F. T. Characterization of messenger RNA populations of Crithidia fasciculata. Arch Biochem Biophys. 1981 Jul;209(2):349–355. doi: 10.1016/0003-9861(81)90291-5. [DOI] [PubMed] [Google Scholar]
- Spencer R., Cross G. A. Purification and properties of nucleic acids from an unusual cytoplasmic organelle in the flagellate protozoan Crithidia oncopelti. Biochim Biophys Acta. 1975 May 1;390(2):141–154. doi: 10.1016/0005-2787(75)90337-8. [DOI] [PubMed] [Google Scholar]
- Stolf A. M., D'Abronzo F. H., Wang L., Salles J. M., Andrade H. F., Jr, Silva S. M., Brentani R. Selection of albumin mRNA by thioacetamide. Biochem Biophys Res Commun. 1976 Oct 18;72(4):1576–1584. doi: 10.1016/s0006-291x(76)80194-5. [DOI] [PubMed] [Google Scholar]
- Tanaka N., Kinoshita T., Masukawa H. Mechanism of protein synthesis inhibition by fusidic acid and related antibiotics. Biochem Biophys Res Commun. 1968 Feb 15;30(3):278–283. doi: 10.1016/0006-291x(68)90447-6. [DOI] [PubMed] [Google Scholar]
- Tanowitz H., Wittner M., Sveda M., Soeiro R. Studies of ribosomal RNA of Trypanosoma cruzi. J Parasitol. 1975 Dec;61(6):1065–1069. [PubMed] [Google Scholar]
- Williams R. O., Marcu K. B., Young J. R., Rovis L., Williams S. C. A characterization of mRNA activites and their sequence complexities in Trypanosoma brucei: partial purification and properties of the VSSA mRNA. Nucleic Acids Res. 1978 Sep;5(9):3171–3182. doi: 10.1093/nar/5.9.3171. [DOI] [PMC free article] [PubMed] [Google Scholar]

