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Immunoreactivity of the two common allozymes of murine glucosephosphate isomerase

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Abstract

There are two common electrophoretic variants (allozymes) of murine glucosephosphate isomerase (GPI). In order to develop immunocytochemical procedures that are allozyme-specific, the two variants (GPI-1A and GPI-1B) were purified from skeletal muscle of several mouse strains and used as antigens for the elicitation of antibodies. The allozymes were purified to a specific activity of 800 units/mg by substrate elution from cellulose phosphate. When the purified allozymes were presented as antigens to goats, rabbits, and mice (both syngeneically and allogeneically), the goats and rabbits produced high titers of anti-GPI antibody, but no humoral antibody was detected in the mice, as determined by radioimmunoassays. Antisera specific for the GPI-1B variant were enriched by absorbing selected sera with GPI-1A conjugated to Sepharose 4B. No antisera specific for GPI-1A were detected following the immunizations. The specificity of the anti-GPI-1B antisera provides a unique opportunity for the development of immunocytochemical procedures for studying the distribution of this allozyme in chimeric mouse tissues.

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References

  • Baughan, M., Valentine, W., Paglia, D., Ways, P., Simon, E., and deMarsh Q. (1968). Hereditary hemolytic anemia associated with glucosephosphate isomerase (GPI) deficiency—a new enzyme defect of human erythrocytes. Blood 32236.

    Google Scholar 

  • Bosma, M. J., Marks, R., and DeWitt, C. L. (1975). Quantitation of mouse immunoglobulin allotypes by a modified solid-phase radioimmune assay. J. Immunol. 1151381.

    Google Scholar 

  • Carter, N. D., and Parr, C. W. (1967). Isoenzymes of phosphoglucose isomerase in mice. Nature 216511.

    Google Scholar 

  • Charles, D. J., and Lee, C.-Y. (1980). Biochemical and immunological characterization of genetic variants in phosphoglucose isomerase from mouse. Biochem. Genet. 18153.

    Google Scholar 

  • DeLorenzo, R. J., and Ruddle, F. H. (1969). Genetic control of two electrophoretic variants of glucosephosphate isomerase in the mouse (Mus musculus). Biochem. Genet. 3151.

    Google Scholar 

  • Gearhart, J., and Mintz, B. (1972). Clonal origins of somites and their muscle derivatives: evidence from allophenic mice. Dev. Biol. 2927.

    Google Scholar 

  • Gearhart, J., and Oster-Granite, M. L. (1978). Antibodies to allozymes as potential cell markers for chimeric mouse studies. In Russell, L. B. (ed.), Genetic Mosaics and Chimeras in Mammals Plenum Press, New York, pp. 111–122.

    Google Scholar 

  • Gearhart, J., and Oster-Granite, M. L. (1980). An immunofluorescence procedure for the tissue localization of glucosephosphate isomerase. J. Histochem. Cytochem. 28245.

    Google Scholar 

  • Gearhart, J., and Oster-Granite, M. L. (1981). Distribution of glucosephosphate isomerase in maturing oocytes and early embryonic stages of the mouse. Biol. Reprod. 23837.

    Google Scholar 

  • Gibson, D. R., Talent, J. M., Gracy, R. W., Schnackerz, K. D., and Ishimoto, G. (1978). A preparative method for the isolation of genetic variant forms of glucosephosphate isomerase and a study of five variants. Clin. Chim. Acta 89355.

    Google Scholar 

  • Grabar, P., and Williams, Jr., C. A., (1953). Methode permettant l'etude conjugee des proprietes electrophoretiques et immunochimiques d'un melange de proteines. Application au serum sanguin. Biochim. Biophys. Acta 10193.

    Google Scholar 

  • Gracy, R. W., and Tilley, B. E. (1975). Phosphoglucose isomerase of human erythrocytes and cardiac tissue. In Colowick, S. P., and Kaplan, N. O. (eds.), Methods in Enzymology, Vol. 41, Academic Press, New York, pp. 392–400.

    Google Scholar 

  • Hunter, W. M., and Greenwood, F. C. (1962). Preparation of iodine-131 labeled human growth hormone of high specific activity. Nature 194495.

    Google Scholar 

  • Hutton, J., and Chilcote, R. (1974). Glucosephosphate isomerase deficiency with hereditary nonspherocytic hemolytic anemia. J. Pediatr. 85494.

    Google Scholar 

  • King, J., and Laemmli, U. (1971). Polypeptides of the tail fibers of bacteriophage T4. J. Mol. Biol. 62465.

    Google Scholar 

  • Lowry, O. H., Rosenbrough, N. J., Farr, A. L., and Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193265.

    Google Scholar 

  • McLaren, A. (1976). Mammalian Chimaeras Cambridge University Press, New York.

    Google Scholar 

  • Noltmann, E. A. (1966). Phosphoglucose isomerase. I. Rabbit muscle (crystalline). In Wood, W. A. (ed.), Methods in Enzymology, Vol. 9, Academic Press, New York, pp. 557–565.

    Google Scholar 

  • Noltmann, E. A. (1975). The different origin of multiple molecular forms of phosphoglucose isomerase. In Markert, C. L. (ed.), Isozymes, Vol. I, Academic Press, New York, pp. 451–470.

    Google Scholar 

  • Oster-Granite, M. L., and Gearhart, J. (1980). Immunofluorescence and histochemical localization of glucosephosphate isomerase in neural tissues. J. Histochem. Cytochem. 28250.

    Google Scholar 

  • Oster-Granite, M. L., and Gearhart, J. (1981a). Cell lineage analyses of Purkinje cells in murine chimeras. In Palay, S., and Chan-Palay, V. (eds.), Cerebellum-New Vistas Springer-Verlag, New York (in press).

    Google Scholar 

  • Oster-Granite, M. L., and Gearhart, J. (1981b). Cell lineage analyses of Purkinje cells in mouse chimeras. Dev. Biol. (in press).

  • Padua, R. A., Bulfield, G., and Peters, J. (1978). Biochemical genetics of a new glucosephosphate isomerase allele (Gpi-I c) from wild mice. Biochem Genet. 16127.

    Google Scholar 

  • Paglia, D. E., and Valentine, W. N. (1974). Hereditary glucosephosphate isomerase deficiency. Am. J. Clin. Pathol. 62740.

    Google Scholar 

  • Peterson, E. A., and Sober, H. A. (1962). Column chromatography of proteins: substituted celluloses. In Colowick, S. P., and Kaplan, N. O. (eds.), Methods in Enzymology, Vol. 5, Academic Press, New York, pp. 3–27.

    Google Scholar 

  • Peterson, A., and Wong, G. (1978). Genetic regulation of glucosephosphate isomerase in mouse oocytes. Nature 276267.

    Google Scholar 

  • Phillips, T. L., Talent, J. M., and Gracy, R. W. (1976). Isolation of rabbit muscle glucosephosphate isomerase by a single step substrate elution. Biochim. Biophys. Acta 429624.

    Google Scholar 

  • Righetti, P. G., and Drysdale, J. W. (1976). Isoelectric focusing. In Work, T., and Work, E. (eds.), Laboratory Techniques in Biochemistry and Molecular Biology North-Holland, Amsterdam, pp. 450–469.

    Google Scholar 

  • Swenstrom, R., and Shank, P. R. (1978). X-ray intensifying screens greatly enhance the detection by autoradiography of the radioactive isotopes 32P and 125I. Anal. Biochem. 86184.

    Google Scholar 

  • Tilley, B. E., Gracy, R. W., and Welch, S. G. (1974). A point mutation increasing the stability of phosphoglucose isomerase. J. Biol. Chem. 2494571.

    Google Scholar 

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This research was supported by grants from the Public Health Service (HD 12685) and the National Science Foundation (PCM 7923065). J.G. is the recipient of a Basil O'Connor Award from the March of Dimes-Birth Defects, and M.L.O.G. is a Fellow of the Alfred Sloan Foundation.

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Gearhart, J., Oster-Granite, M.L. & Musser, J.M. Immunoreactivity of the two common allozymes of murine glucosephosphate isomerase. Biochem Genet 19, 445–455 (1981). https://doi.org/10.1007/BF00484617

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  • DOI: https://doi.org/10.1007/BF00484617

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