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Detection of serum proteins in the electrophoretic patterns of total proteins of mycoplasma cells

Published online by Cambridge University Press:  15 May 2009

O. E. Yaguzhinskaya
O. E. Yaguzhinskaya
Affiliation:
Institute of Chemical Physics, Academy of Sciences, Moscow, U.S.S.R
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The contamination of mycoplasma cell preparations by serum proteins originating from culture medium was studied. A. laidlawii and M. arthritidis cells were grown in the presence of [14C]-aminoacids, and the cells were washed with 0·9% NaCl by threefold centrifugation. Total proteins of the washed cells were analysed by SDS gel electrophoresis. Coomassie-stained electrophoretic patterns were compared with autoradiographs of the same gels. The stained electrophoretic pattern of washed A. laidlawii grown without serum was identical with autoradiographs of the same cells grown without or with serum. That of washed A. laidlawii grown with serum differed from the corresponding autoradiograph by the presence of extra protein bands I, II, III, and IV with molecular weights of over 160,000, 80,000–87,000, 55,000 and 25,000, respectively. The same extra bands were found in stained electrophoretic patterns of washed: (a) A. laidlawii cells grown without serum and mixed with serum in the stationary phase, (b) M. arthritidis cells, as compared with their autoradiographs, (c) serum precipitate. The bands III and IV may be due to the heavy and light chains of γ-globulin, the band II might belong to transferrin or to some component of complement. Acidification of serum to pH 5 brought about 100-fold rise of amount of serum precipitate, the number of bands in the electrophoretic pattern of the precipitate being also increased. Stained electrophoretic patterns of cells purified by twofold centrifugation in step sucrose density gradient (1·20–1·27 g./cm.3 for A. laidlawii, and 1·15–1·25 for M. arthritidis) contained no extra bands and matched completely with their autoradiographs.

It was concluded that contamination of washed mycoplasma cells by serum proteins is mainly due to co-precipitation of aggregated serum proteins together with cells during centrifugation rather than to adsorption of serum proteins on the cell surface.

Type
Research Article
Information
Journal of Hygiene , Volume 77 , Issue 2 , October 1976 , pp. 189 - 198
Copyright
Copyright © Cambridge University Press 1976

References

REFERENCES

Andreev, V. M. & Gonikberg, E. M. (1974). New postsynthetic modifications of proteins in the assembly of bacteriophage T4 head. Doklady Akademii Nauk SSSR 218, 222. (In Russian.)Google Scholar
Asmar, J. A. (1965). Chromatographic and immunoelectrophoretic characterization of chicken and turkey agglutinating antibodies to Mycoplasma gallisepticum. Ph.D. Thesis. University of California. (Cited by Bradbury & Jordan, 1972.)Google Scholar
Bradbury, J. M. & Jordan, F. T. W. (1971). The influence of pH of the culture medium on the sensitivity of Mycoplasma gallisepticum antigens for use in certain serological tests. Journal of Hygiene 69, 593.Google ScholarPubMed
Bradbury, J. M. & Jordan, F. T. W. (1972). Studies on adsorption of certain medium proteins to Mycoplasma gallisepticum and their influence on agglutination and haemagglutination reactions. Journal of Hygiene 70, 267.CrossRefGoogle ScholarPubMed
Chelton, E. T., Jones, A. S. & Walker, R. T. (1968). The chemical composition of nucleic acids and the proteins of some Mycoplasma strains. Journal of General Microbiology 50, 305.Google Scholar
Ishizaka, T., Ishizaka, K. & Borsos, T. (1961). Biological activity of aggregated γ-globulin. Journal of Immunology 87, 433.CrossRefGoogle Scholar
Jones, A. S., Terriennson, J. R. & Walker, R. T. (1965). The chemical composition of nucleic acids and other macromolecular constituents of Mycoplasma mycoides var. capri. Journal of General Microbiology 40, 405.CrossRefGoogle ScholarPubMed
King, J. & Laemmli, U. K. (1971). Polypeptides of tail fibres of bacteriophage T4. Journal of Molecular Biology 62, 465.Google Scholar
King, J. & Laemmli, U. K. (1973). Bacteriophage T4 tail assembly: structural proteins and their genetic identification. Journal of Molecular Biology 75, 315.CrossRefGoogle ScholarPubMed
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London, 227, 680.CrossRefGoogle ScholarPubMed
Lipman, P. R., Clyde, W. A. J. R. & Denny, F. W. (1969). Characteristics of virulent, attenuated, and avirulent Mycoplasma pneumoniae strains. Journal of Bacteriology 100, 1037.Google Scholar
Lomonosov, J. J. & Soshin, L. D. (1968). Measurement of tritium, p. 77. Moskow, Atomisdat. (In Russian.)Google Scholar
Lozinsky, T. F., Gushchin, B. V., Astakhova, A. V., Klimenko, S. M. & Zhdanov, V. M. (1974). Detection of latent viruses and mycoplasmas by molecular biology methods. Voprosy virusologii No 4, 474. (In Russian.)Google Scholar
Palmour, R. M. & Stutton, H. E. (1971). Vertebrate transferrin. Molecular weights, chemical composition, and iron-binding studies. Biochemistry 10, 4026.CrossRefGoogle Scholar
Pollack, J. D., Sommerson, N. L. & Senterfit, L. B. (1969). Effect of pH on the immunogenicity of Mycoplasma pneumonias. Journal of Bacteriology 97, 612.CrossRefGoogle Scholar
Rottem, S., Hasin, M. & Razin, S. (1973). Binding of proteins to mycoplasma membranes Biochimica et biophysica acta 298, 876.CrossRefGoogle ScholarPubMed
Sethi, K. K. & Brandis, H. (1972). Killing of mycoplasmas by the antibodies to foreign antigens acquired by the organisms from the growth medium. Medical Microbiology and Immunology 157, 113.CrossRefGoogle ScholarPubMed
Smirnova, T. D., Yaguzhinskaya, O. E. & Balaeva, E. Ya. (1975). Identification of mycoplasma isolated from monkeys. Zhurnal microbiologii, No 1, 90. (In Russian.)Google Scholar
Todaro, G. J., Aaronson, S. A. & Rands, E. (1971). Rapid detection of mycoplasma-infected cell cultures. Experimental Cell Research 65, 256.Google Scholar
Tukachinsky, S. E. & Badyina, V. M. (1974). Inhibition of the anti-complementary activity of gamma-globulin on addition of transferrin. Zhurnal microbiologii No 6, 107. (In Russian.)Google Scholar
Ward, S. (1970). An improved transverse destaining apparatus for acrylamide gels. Analytical Biochemistry 33, 259.CrossRefGoogle ScholarPubMed
Weber, K. & Osborn, M. (1969). The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. Journal of Biological Chemistry 244, 406.CrossRefGoogle ScholarPubMed
Yaguzhinskaya, O. E., Rakovskaya, I. V., Smirnova, T. D., Grebnev, G. M. & Kagan, G. Ya. (1974). Identification of mycoplasma and L-forms of bacteria by electrophoresis in polyacrylamide gel. Zhurnal Microbiologii No 4, 85. (In Russian.)Google Scholar