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. 1976 Sep;73(9):3288–3292. doi: 10.1073/pnas.73.9.3288

Inhibition of erythrocyte sickling by cystamine, a thiol reagent.

W Hassan, Y Beuzard, J Rosa
PMCID: PMC431012  PMID: 135260

Abstract

Incubation of sickle cells with cystamine, a thiol reagnet, resulted in the formation of an intracellular S-ethylamine derivative. The rate of the reaction was dependent upon the cystamine concentration, the temperature, and the duration of the incubation. The cystamine-treated cells demonstrated a marked inhibition of sickling under hypoxic conditions, a decrease in their mean corpuscular hemoglobin concentration, and a significant increase in their oxygen affinity. The oxygen affinity of these cells was less dependent on their mean corpuscular hemoglobin concentration than that of untreated sickle cells. The minimum gelling concentration of S-ethylamine doxyhemoglobin S was slightly increased. Cystamine did not affect the intracellular pH nor the 2,3-diphosphoglycerate level. The exact contribution of the interrelated factors in cystamine inhibition of sickling (changes in oxygen affinity, mean corpuscular hemoglobin concentration, and minimum gelling concentration) has yet to be determined.

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Selected References

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  1. ALLEN D. W., WYMAN J., Jr Equilibre de l'hémoglobine de drépanocytose avec l'oxygène. Rev Hematol. 1954;9(2):155–157. [PubMed] [Google Scholar]
  2. BENESCH R., MACDUFF G., BENESCH R. E. DETERMINATION OF OXYGEN EQUILIBRIA WITH A VERSATILE NEW TONOMETER. Anal Biochem. 1965 Apr;11:81–87. doi: 10.1016/0003-2697(65)90045-x. [DOI] [PubMed] [Google Scholar]
  3. BESSIS M., NOMARSKI G., THIERY J. P., BRETON-GORIUS J. Etude sur la falciformation des globules rouges au microscope polarisant et au microscope électronique. II. L'intérieru du globule; comparaison avec les cristaux intra-globulaires. Rev Hematol. 1958 Apr-Jun;13(2):249–270. [PubMed] [Google Scholar]
  4. Bellingham A. J., Huehns E. R. Compensation in haemolytic anaemias caused by abnormal haemoglobins. Nature. 1968 Jun 8;218(5145):924–926. doi: 10.1038/218924a0. [DOI] [PubMed] [Google Scholar]
  5. Benesch R. E., Benesch R., Yung S. Equations for the spectrophotometric analysis of hemoglobin mixtures. Anal Biochem. 1973 Sep;55(1):245–248. doi: 10.1016/0003-2697(73)90309-6. [DOI] [PubMed] [Google Scholar]
  6. Bookchin R. M., Nagel R. L. Conformational requirements for the polymerization of hemoglobin S: studies of mixed liganded hybrids. J Mol Biol. 1973 May 15;76(2):233–239. doi: 10.1016/0022-2836(73)90387-2. [DOI] [PubMed] [Google Scholar]
  7. Bookchin R. M., Nagel R. L. Ligand-induced conformational dependence of hemoglobin in sickling interactios. J Mol Biol. 1971 Sep 14;60(2):263–270. doi: 10.1016/0022-2836(71)90292-0. [DOI] [PubMed] [Google Scholar]
  8. Cerami A., Manning J. M. Potassium cyanate as an inhibitor of the sickling of erythrocytes in vitro. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1180–1183. doi: 10.1073/pnas.68.6.1180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Elbaum D., Nagel R. L., Bookchin R. M., Herskovits T. T. Effect of alkylureas on the polymerization of hemoglobin S. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4718–4722. doi: 10.1073/pnas.71.12.4718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Finch J. T., Perutz M. F., Bertles J. F., Döbler J. Structure of sickled erythrocytes and of sickle-cell hemoglobin fibers. Proc Natl Acad Sci U S A. 1973 Mar;70(3):718–722. doi: 10.1073/pnas.70.3.718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Garel M. C., Cohen-Solal M., Blouquit Y., Rosa J. A method for isolation of abnormal haemoglobins with high oxygen affinity due to a frozen quaternary r-structure: application to Hb Creteil alpha 2 A beta 2 (F5) 89 ASN. FEBS Lett. 1974 Jul 1;43(1):93–96. doi: 10.1016/0014-5793(74)81113-0. [DOI] [PubMed] [Google Scholar]
  12. Garel M. C., Hassan W., Coquelet M. T., Goossens M., Rosa J., Arous N. Hemoglobin J Cairo: beta 65 (E9) Lys leads to Gln, A new hemoglobin variant discovered in an Egyptian family. Biochim Biophys Acta. 1976 Jan 20;420(1):97–104. doi: 10.1016/0005-2795(76)90348-2. [DOI] [PubMed] [Google Scholar]
  13. Hofrichter J., Ross P. D., Eaton W. A. Kinetics and mechanism of deoxyhemoglobin S gelation: a new approach to understanding sickle cell disease. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4864–4868. doi: 10.1073/pnas.71.12.4864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jensen M., Bunn H. F., Halikas G., Kan Y. W., Nathan D. G. Effects of cyanate and 2,3-diphosphoglycerate on sickling. Relationship to oxygenation. J Clin Invest. 1973 Oct;52(10):2542–2547. doi: 10.1172/JCI107445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lubin B. H., Pena V., Mentzer W. C., Bymun E., Bradley T. B., Packer L. Dimethyl adipimidate: a new antisickling agent. Proc Natl Acad Sci U S A. 1975 Jan;72(1):43–46. doi: 10.1073/pnas.72.1.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. MURAYAMA M. Titratable sulfhydryl groups of normal and sickle cell hemoglobins at O degrees and 38 degrees. J Biol Chem. 1957 Sep;228(1):231–240. [PubMed] [Google Scholar]
  17. May A., Huehns E. R. The concentration dependence of the oxygen affinity of haemoglobin S. Br J Haematol. 1975 Jul;30(3):317–335. doi: 10.1111/j.1365-2141.1975.tb00548.x. [DOI] [PubMed] [Google Scholar]
  18. May A., Huehns E. R. The mechanism of the low oxygen affinity of red cells in sickle cell disease. Hamatol Bluttransfus. 1972;10:279–283. [PubMed] [Google Scholar]
  19. Mordukhovich V. V. Soderzhanie beta-merkaptoetilamina v tkaniiakh myshei pri sovmestnom primenenii tsistamina i 5-metoksitriptamina. Radiobiologiia. 1975 Jan-Feb;15(1):132–136. [PubMed] [Google Scholar]
  20. Perutz M. F., Fersht A. R., Simon S. R., Roberts G. C. Influence of globin structure on the state of the heme. II. Allosteric transitions in methemoglobin. Biochemistry. 1974 May 7;13(10):2174–2186. doi: 10.1021/bi00707a027. [DOI] [PubMed] [Google Scholar]
  21. Rose Z. B., Liebowitz J. Direct determination of 2,3-diphosphoglycerate. Anal Biochem. 1970 May;35(1):177–180. doi: 10.1016/0003-2697(70)90023-0. [DOI] [PubMed] [Google Scholar]
  22. SINGER K., SINGER L. Studies on abnormal hemoglobins. VIII. The gelling phenomenon of sickle cell hemoglobin: its biologic and diagnostic significance. Blood. 1953 Nov;8(11):1008–1023. [PubMed] [Google Scholar]
  23. Seakins M., Gibbs W. N., Milner P. F., Bertles J. F. Erythrocyte Hb-S concentration. An important factor in the low oxygen affinity of blood in sickle cell anemia. J Clin Invest. 1973 Feb;52(2):422–432. doi: 10.1172/JCI107199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Smithies O. Disulfide-bond cleavage and formation in proteins. Science. 1965 Dec 17;150(3703):1595–1598. doi: 10.1126/science.150.3703.1595. [DOI] [PubMed] [Google Scholar]
  25. Taylor J. F., Antonini E., Brunori M., Wyman J. Studies on human hemoglobin treated with various sulfhydryl reagents. J Biol Chem. 1966 Jan 10;241(1):241–248. [PubMed] [Google Scholar]
  26. Vasin M. V., Lebedeva N. N. Vliianie tsistamina na rabotosposobnost' chelovaka-operatora. Kosm Biol Aviakosm Med. 1975 Sep-Oct;9(5):54–57. [PubMed] [Google Scholar]
  27. Zak S. J., Geller G. R., Finkel B., Tukey D. P., McCormack M. K., Krivit W. Bis-(N-maleimidomethyl) ether: an antisickling reagent. Proc Natl Acad Sci U S A. 1975 Oct;72(10):4153–4156. doi: 10.1073/pnas.72.10.4153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Zarkowsky H. S., Hochmuth R. M. Sickling times of individual erythrocytes at zero Po2. J Clin Invest. 1975 Oct;56(4):1023–1034. doi: 10.1172/JCI108149. [DOI] [PMC free article] [PubMed] [Google Scholar]

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