Humangenetik

, Volume 10, Issue 3, pp 224–230 | Cite as

Detection of phosphohexose isomerase deficiency in human fibroblast cultures

  • W. Krone
  • G. Schneider
  • D. Schulz
  • H. Arnold
  • K. G. Blume
Original Investigations

Summary

Fibroblasts cultured from two patients afflicted with nonspherocytic hemolytic anemia due to phosphohexose isomerase (PHI) deficiency show on the average 53% of the normal PHI-activity. The presence of the defective enzyme in cells derived from the heterozygous relatives of the patients is revealed by an intermediate average specific activity; the wide range of PHI-activities observed in these cells, however, precludes the detection of heteozygotes. The PHI-genotypes of the patients and of their heterozygous and normal relatives respectively, can be distinguished by starch gel electrophoresis and by heat-inactivation studies with fibroblast-homogenates. These latter experiments confirm the results obtained with hemolysates (Tariverdian et al., 1970).

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Arnold, H., Blume, K. G., Busch, D., Lenkeit, U., Löhr, G. W., Lübs, E.: Klinische und biochemische Untersuchungen zur Glucosephosphatisomerase normaler menschlicher Erythrocyten und bei Glucosephosphatisomerase-Mangel. Klin. Wschr., in press (1970).Google Scholar
  2. Baughan, M. A., Valentine, W. N., Paglia, D. E., Ways, P. O., Simons, E. R., DeMarsh, A. B.: Hereditary hemolytic anemia associated with glucosephosphate isomerase (GPI) deficiency—a new enzyme defect of human erythrocytes. Blood 32, 236–249 (1968).Google Scholar
  3. Davidson, R. G.: Application of cell culture techniques to human genetics, in: Modern trends in human genetics, Emery, A.E.H., editor; vol. 1, p. 143–180. London: Butterworth 1970.Google Scholar
  4. DeMars, R.: Some studies of enzymes in cultivated human cells. Nat. Cancer Inst. Monogr. 13, 181–193 (1964).Google Scholar
  5. Detter, J. C., Ways, P. O., Giblett, E. R., Baughan, M. A., Hopkinson, D. A., Povey, S., Harris, H.: Inherited variation in human phosphohexose isomerase. Ann. hum. Genet. 31, 329–338 (1968).Google Scholar
  6. Evans, V. J., Earle, W. R.: The use of perforated cellophan for the growth of cells in tissue culture. J. nat. Cancer Inst. 8, 103 (1947).Google Scholar
  7. Lowry, O. H., Roseburough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin phenol reagent. J. biol. Chem. 193, 265–275 (1959).Google Scholar
  8. Paglia, D. E., Holland, P., Baughan, M. A., Valentine, W. N.: Occurrence of defective hexosephosphate isomerization in human erythrocytes and leukocytes. New. Engl. J. Med. 280, 66–71 (1969).Google Scholar
  9. Schimke, R. T.: On the roles of synthesis and degradation in regulation of enzyme levels in mammalian tissues, in: Current topics in cellular regulation, Horecker, B. L., Stadtman, E. R., editors; vol. 1, p. 77–124. New York-London: Academic Press 1969.Google Scholar
  10. Selvyn, J. G., Dacie, J. V.: Autohemolysis and other changes resulting from the incubation in vitro of red cells from patients with congenital hemolytic anemia. Blood 9, 414 (1954).Google Scholar
  11. Tariverdian, G., Arnold, H., Blume, K. G., Lenkeit, U., Löhr, G. W.: Zur Formalgenetik der Phosphoglucoseisomerase (E.C.: 5.3.1.9); Untersuchung einer Sippe mit PGI-Defizienz. Humangenetik 10, 218–223 (1970).Google Scholar

Copyright information

© Springer-Verlag 1970

Authors and Affiliations

  • W. Krone
    • 1
  • G. Schneider
    • 1
  • D. Schulz
    • 1
  • H. Arnold
    • 2
  • K. G. Blume
    • 2
  1. 1.Institut für Humangenetik und Anthropologie Freiburg i. Br.Freiburg i. Br.Deutschland
  2. 2.Medizinische Universitätsklinik Freiburg i. B.Freiburg i. B.Deutschland

Personalised recommendations