Abstract
The covalent fixation of benzenehexacarboxylate (BHC) onto dextran was carried out according to several reaction schemes. The polyanionic polymers thus synthesized were capable of decreasing the oxygen affinity of hemoglobin by specifically interacting with the 2,3-diphosphoglycerate (2,3-DPG) binding site of the protein. The intensity of this effect was correlated to both the chemical structure of the polyanionic polymers and the BHC content in polymer. The polyanionic polymer, containing 0.035 mol BHC/g and presenting no cross-linking between its polymer chains, possessed the best effector properties. These properties were used to direct the covalent fixation of the dextran-benzenehexacarboxylate onto the phosphate binding site of the protein. The resulting hemoglobin was mainly substituted at the same time by one or more linked BHC onto bothαΒ dimers in the vicinity of the 2,3-DPG site. Thus, the modification of hemoglobin led to an increase in the hydrodynamic volume of each dimer sufficient to limit the diffusion of the conjugates through the kidney membrane, even if the conjugates had dissociated intoαΒ dimers. Compared to that of free hemoglobin, the oxygen affinity of the conjugates was significantly decreased. This type of covalent conjugate exhibited general properties quite suitable for use as blood substitutes.
Similar content being viewed by others
Abbreviations
- Hb:
-
hemoglobin
- 2,3-DPG:
-
2,3-diphosphoglycerate
- BHC:
-
benzenehexacarboxylate
- IHP:
-
inositolhexaphosphate
- EDCI:
-
3-ethyl-1-(3-dimethylaminopropyl) carbodiimide hydrochloride
References
Arnone, A., and Perutz M. F. (1974).Nature 249, 35–36.
Benesch, R., Benesch, R. E., Kwong, S., Acharya, A. S., and Manning, J. M. (1982).J. Biol. Chem. 257, 1320–1324.
Benesch, R. E., and Kwong, S. (1988).Biochem. Biophys. Res. Comm. 158, 9–14.
Benesch, R. E., and Kwong, S. (1991).J. Prot. Chem. 10, 503–510.
Bucci, E. (1974).Biochemistry 13, 814–820.
Dellacherie, E., Bonneaux, F., Labrude, P., and Vigneron C. (1983).Biochem. Biophys. Acta 749, 106–114.
Desbos, A., and Banerjee, R. (1975).J. Mol. Biol. 92, 479–493.
Evelyn, K. A., and Malloy, H. T. (1938).J. Biol. Chem. 126, 655–662.
Gray, D. R., and Gibson, Q. H. (1971).J. Biol. Chem. 246, 7168–7174.
Kaplan, J. C. (1965).Rev. FranÇ. Etudes Clin. Biol. 10, 856–859.
Kavanaugh, M. P., Shih, D. T. B., and Jones, R. T. (1988).Biochemistry 27, 1804–1808.
Prouchayret, F., Sacco, D., and Dellacherie, E. (1989).Polym. Bull. 21, 309–313.
Sacco, D., Bonneaux, F., and Dellacherie, E. (1988).Int. J. Biol. Maromol. 10, 305–310.
Sacco, D., Prouchayret, F., Dellacherie, E. (1989).Makromol. Chem. 190, 1671–1680.
Synder, S. R., Welty, E. V., Walder, R. Y., Williams, L. A., and Walder, J. A. (1987).Proc. Natl. Acad. Sci. USA 84, 7280–7284.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sacco, D., Dellacherie, E. & Prouchayret, F. Effect of covalent labeling of dextran-benzenehexacarboxylate on hemoglobin. J Protein Chem 13, 1–8 (1994). https://doi.org/10.1007/BF01891986
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01891986