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. 1981 Apr;146(1):222–232. doi: 10.1128/jb.146.1.222-232.1981

Analysis of multiple isoenzyme expression among twenty-two species of Mycoplasma and Acholeplasma.

S J O'Brien, J M Simonson, M W Grabowski, M F Barile
PMCID: PMC217073  PMID: 7217001

Abstract

Crude extracts of triple-cloned, purified cultures of 22 species of Mycoplasma and Acholeplasma were examined for expression of 21 isozyme systems routinely used to type mammalian cells. Nine previously described enzymes (purine nucleoside phosphorylase, adenylate kinase, dipeptidase, esterase, glyceraldehyde-3-phosphate dehydrogenase, glucose phosphate isomerase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and superoxide dismutase) and three enzymes not previously reported in mycoplasma (triose phosphate isomerase, inorganic pyrophosphatase, and acid phosphatase) were detected in some or all of the species examined. These findings provide new information on the enzymatic expressions of these organisms. Three of the isozyme systems (superoxide dismutase, glucose-6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase) were present in Acholeplasma species but not in any Mycoplasma species. The characteristic pattern of electrophoretic mobility of the 12 isozyme systems also provides a useful biochemical property for identification, characterization, and classification of these mycoplasmas. Mycoplasma isozyme expression for seven of the enzymes were readily detected in various infected-cell culture lines by using either cell extracts or concentrated cell culture fluids. Mycoplasma-specific enzymes found in infected-cell extracts had the same electrophoretic mobility patterns as enzymes obtained from broth-grown mycoplasmas of the same species. Expression of homologous mammalian enzymes was not detectably altered by infection with mycoplasmas.

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Selected References

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  1. Allsop B. A., Matthews K. P. Studies of lactate dehydrogenase of Mycoplasma mycoides var. mycoides. J Gen Microbiol. 1975 May;88(1):58–64. doi: 10.1099/00221287-88-1-58. [DOI] [PubMed] [Google Scholar]
  2. Cherry J. D., Taylor-Robinson D. Peroxide production by mycoplasmas in chicken tracheal organ cultures. Nature. 1970 Dec 12;228(5276):1099–1100. doi: 10.1038/2281099a0. [DOI] [PubMed] [Google Scholar]
  3. Clark A. F., Farrell D. F., Burke W., Scott C. R. The effect of Mycoplasma contamination on the in vitro assay of pyruvate dehydrogenase activity in cultured fibroblasts. Clin Chim Acta. 1978 Jan 2;82(1-2):119–124. doi: 10.1016/0009-8981(78)90034-7. [DOI] [PubMed] [Google Scholar]
  4. Cohen G., Somerson N. L. Mycoplasma pneumoniae: hydrogen peroxide secretion and its possible role in virulence. Ann N Y Acad Sci. 1967 Jul 28;143(1):85–87. doi: 10.1111/j.1749-6632.1967.tb27648.x. [DOI] [PubMed] [Google Scholar]
  5. Cole B. C., Ward J. R., Martin C. H. Hemolysin and peroxide activity of Mycoplasma species. J Bacteriol. 1968 Jun;95(6):2022–2030. doi: 10.1128/jb.95.6.2022-2030.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fridovich I. The biology of oxygen radicals. Science. 1978 Sep 8;201(4359):875–880. doi: 10.1126/science.210504. [DOI] [PubMed] [Google Scholar]
  7. Hatanaka M., Del Giudice R., Long C. Adenine formation from adenosine by mycoplasmas: adenosine phosphorylase activity. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1401–1405. doi: 10.1073/pnas.72.4.1401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. Lanham S. M., Lemcke R. M., Scott C. M., Grendon J. M. Isoenzymes in two species of Acholeplasma. J Gen Microbiol. 1980 Mar;117(1):19–31. doi: 10.1099/00221287-117-1-19. [DOI] [PubMed] [Google Scholar]
  10. Lavappa K. S., Macy M. L., Shannon J. E. Examination of ATCC stocks for HeLa marker chromosomes in human cell lines. Nature. 1976 Jan 22;259(5540):211–213. doi: 10.1038/259211a0. [DOI] [PubMed] [Google Scholar]
  11. Lemons R. S., Nash W. G., O'Brien S. J., Benveniste R. E., Sherr C. J. A gene (Bevi) on human chromosome 6 is an integration site for baboon type C DNA provirus in human cells. Cell. 1978 Aug;14(4):995–1005. doi: 10.1016/0092-8674(78)90353-7. [DOI] [PubMed] [Google Scholar]
  12. Long C. W., DelGiudice R., Gardella R. S., Hatanaka M. Uracil phosphoribosyl transferase activity of mycoplasma and infected cell cultures. In Vitro. 1977 Jul;13(7):429–433. doi: 10.1007/BF02615103. [DOI] [PubMed] [Google Scholar]
  13. Markert C. L., Whitt G. S. Molecular varieties of isozymes. Experientia. 1968 Oct 15;24(10):977–991. doi: 10.1007/BF02138698. [DOI] [PubMed] [Google Scholar]
  14. Mitchell A., Sin I. L., Finch L. R. Enzymes of purine metabolism in Mycoplasma mycoides subsp. mycoides. J Bacteriol. 1978 Jun;134(3):706–712. doi: 10.1128/jb.134.3.706-712.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nelson-Rees W. A., Flandermeyer R. R. HeLa cultures defined. Science. 1976 Jan 9;191(4222):96–98. doi: 10.1126/science.1246601. [DOI] [PubMed] [Google Scholar]
  16. Nichols E. A., Ruddle F. H. A review of enzyme polymorphism, linkage and electrophoretic conditions for mouse and somatic cell hybrids in starch gels. J Histochem Cytochem. 1973 Dec;21(12):1066–1081. doi: 10.1177/21.12.1066. [DOI] [PubMed] [Google Scholar]
  17. Nichols E. A., Ruddle F. H. Comparative sensitivities of electrophoretic assays for human enzymes. Biochem Genet. 1979 Feb;17(1-2):127–132. doi: 10.1007/BF00484478. [DOI] [PubMed] [Google Scholar]
  18. O'Brien S. J. Bvr-1, a restriction locus of a type C RNA virus in the feline cellular genome: identification, location, and phenotypic characterization in cat X mouse somatic cell hybrids. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4618–4622. doi: 10.1073/pnas.73.12.4618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. O'Brien S. J., Shannon J. E., Gail M. H. A molecular approach to the identification and individualization of human and animal cells in culture: isozyme and allozyme genetic signatures. In Vitro. 1980 Feb;16(2):119–135. doi: 10.1007/BF02831503. [DOI] [PubMed] [Google Scholar]
  20. O'Brien S. U., Kleiner G., Olson R., Shannon J. E. Enzyme polymorphisms as genetic signatures in human cell cultures. Science. 1977 Mar 25;195(4284):1345–1348. doi: 10.1126/science.841332. [DOI] [PubMed] [Google Scholar]
  21. Pollack J. D., Razin S., Cleverdon R. C. Localization of Enzymes in Mycoplasma. J Bacteriol. 1965 Sep;90(3):617–622. doi: 10.1128/jb.90.3.617-622.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Razin S., Rottem S. Identification of Mycoplasma and other microorganisms by polyacrylamide-gel electrophoresis of cell proteins. J Bacteriol. 1967 Dec;94(6):1807–1810. doi: 10.1128/jb.94.6.1807-1810.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rose N. R., Milisauskas V., Kite J. H., Jr Effect of mycoplasma infection on esterase activity of human FL amnion and WI-38 cells. Proc Soc Exp Biol Med. 1972 Jun;140(2):391–394. doi: 10.3181/00379727-140-36465. [DOI] [PubMed] [Google Scholar]
  24. Stanbridge E. J., Tischfield J. A., Schneider E. L. Appearance of hypoxanthine guanine phosphoribosyltransferase activity as a consequence of mycoplasma contamination. Nature. 1975 Jul 24;256(5515):329–331. doi: 10.1038/256329a0. [DOI] [PubMed] [Google Scholar]
  25. Thomas M. A., Shipman C., Jr, Sandberg J. N., Drach J. C. Adenosine phosphorylase activity in a mutant HEp-2 cell line contaminated with Mycoplasm hyorhinis. In Vitro. 1977 Aug;13(8):502–509. doi: 10.1007/BF02615143. [DOI] [PubMed] [Google Scholar]
  26. Uitendaal M. P., DeBruyn C. H., Hatanaka M., Hösli P. An ultramicrochemical test for mycoplasmal contamination of cultured cells. In Vitro. 1979 Feb;15(2):103–108. doi: 10.1007/BF02618104. [DOI] [PubMed] [Google Scholar]
  27. Van Roy F., Fiers W. Mycoplasma in African green monkey kidney cell cultures: biochemical detection and effects in virus-infected cells. In Vitro. 1977 Jun;13(6):357–365. doi: 10.1007/BF02615095. [DOI] [PubMed] [Google Scholar]

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