Abstract
Kinetic studies on solutions of purified haemoglobin S indicate that the rate of intracellular polymerization is an important variable in the pathophysiology of sickle cell disease1–4. Until now, however, no experimental technique has been available to measure directly the kinetics of intracellular polymerization. Indirect methods, which use visual determination of cellular shape changes or changes in filterability of red cell suspensions, have given apparently conflicting results5–9. Here we report our initial results on the application of a laser-photolysis, light scattering technique to measure directly the kinetics of haemoglobin S polymerization in single red cells. In our experiment, deoxyhaemoglobin S is rapidly formed by photolysing the carbon monoxide complex with an argon ion laser focused inside the cell, and the change in scattered light is used to detect the appearance of polymer. We find a very wide distribution of delay times, ranging from 1 ms to >100s, indicating that the polymerization inside red cells proceeds by the same nucleation and growth mechanism as in solutions of purified haemoglobin S10.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Hofrichter, J., Ross, P. D. & Eaton, W. A. Proc. natn. Acad. Sci. U.S.A. 71, 4864–4868 (1974).
Eaton, W. A., Hofrichter, J. & Ross, P. D. Blood 47, 621–627 (1976).
Sunshine, H. R., Hofrichter, J. & Eaton, W. A. Nature 275, 238–240 (1978).
Sunshine, H. R., Ferrone, F. A., Hofrichter, J. & Eaton, W. A. INSERM Symp. 9, 31–46 (1979).
Rampling, M. W. & Sirs, J. A. Clin. Sci. molec. Med. 45, 655–664 (1973).
Messer, M. J. & Harris, J. W. J. Lab. clin. Med. 76, 537–547 (1970).
Zarkowsky, H. S. & Hochmuth, R. M. J. clin. Invest. 56, 1023–1034 (1975).
Antonini, E. et al. FEBS Lett. 86 209–212 (1978).
Harrington, J. P. & Nagel, R. L. J. Lab. clin. Med. 90, 863–872 (1977).
Ferrone, F. A., Hofrichter, J., Sunshine, H. R. & Eaton, W. A. Biophys. J. 32, 361–377 (1980).
Dvorak, J. A. & Stotler, W. F. Expl Cell. Res. 68, 144–148 (1971).
Parpart, A. K. & Hoffman, J. F. J. cell. comp. Physiol. 47, 295–303 (1956).
Padilla, F., Bromberg, P. A. & Jensen, W. N. Blood 41, 653–660 (1973).
Burton, A. L., Anderson, W. L. & Andrews, R. V. Blood 32, 819–822 (1968).
Seakins, M., Gibbs, W. N., Milner, P. F. & Bertles, J. F. J. clin. Invest. 52, 422–432 (1968).
Hofrichter, J., Ross, P. D. & Eaton, W. A. Proc. natn. Acad. Sci. U.S.A. 73, 3035–3039 (1976).
Goldberg, M. A., Husson, M. A. & Bunn, H. F. J. biol. Chem. 252, 3414–3421 (1977).
Swerdlow, P. H. et al. Hemoglobin 1, 527–537 (1977).
Bookchin, R. M., Ueda, Y., Nagel, R. L. & Landau, L. C. in Biochemical and Clinical Aspects of Hemoglobin Abnormalities (ed. Caughey, W. S.) 57–64 (Academic, New York, 1978).
Serjeant, G. R. The clinical Features of Sickle Cell Disease (North-Holland, Amsterdam, 1974).
Wrightstone, R. N. & Huisman, T. H. J. Am. J. clin. Path. 61, 375–381 (1974).
Powars, D. R., Schroeder, W. A., Weiss, J. N., Chan, L. S. & Azen, S. P. J. clin. Invest. 65, 732–740 (1980).
Fabry, M. E. & Nagel, R. L. Blood Cells 8, 9–15 (1982).
Dover, G. J., Boyer, S. H., Charache, S. & Heintzelman, K. New Engl. J. Med. 299, 1428–1434 (1978).
Sunshine, H. R., Hofrichter, J. & Eaton, W. A. J. molec. Biol. 133, 435–467 (1979).
Goldberg, M. A., Lalos, A. T. & Bunn, H. F. J. biol. Chem. 256, 193–197 (1981).
Bookchin, R. M., Balazs, T. & Landau, L. C. J. Lab. clin. Med. 87, 597–616 (1976).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Coletta, M., Hofrichter, J., Ferrone, F. et al. Kinetics of sickle haemoglobin polymerization in single red cells. Nature 300, 194–197 (1982). https://doi.org/10.1038/300194a0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/300194a0
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.