Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1981 Oct 1;199(1):61–67. doi: 10.1042/bj1990061

Modification by simetryn sulphoxide of a specific thiol group in rat haemoglobin.

G J Hughes, C de Jong, R W Fischer, K H Winterhalter, K J Wilson
PMCID: PMC1163334  PMID: 7337714

Abstract

Native rat haemoglobins were found to bind simetryn sulphoxide to an extent 40-fold greater than human haemoglobin. This specific behaviour was studied by using only high-pressure ('performance') liquid chromatography for the preparative separation of globin chains and the isolation of peptides resulting from chemical and enzymic degradation. High recoveries (greater than 80%) of peptides throughout the procedures in combination with microsequence techniques, allow a definitive assignment of the residue undergoing modification. The haemoglobin beta-chain cystine-125 residue, with a stoichiometry of one per tetramer of rat haemoglobin, was found to be modified. Stereochemical implications of this finding are discussed. Simetryn sulphoxide would appear to be useful as a specific reagent for the mapping of exposed thiol residues in proteins.

Full text

PDF
64

Images in this article

63Fig. 1.
on p.63
64Fig. 2.
on p.64
65Fig. 3.
on p.65
66Fig. 4.
on p.66

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Chua C. G., Carrell R. W., Howard B. H. The amino acid sequence of the alpha chain of the major haemoglobin of the rat (Rattus norvegicus). Biochem J. 1975 Jul;149(1):259–269. doi: 10.1042/bj1490259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chua C. G., Carrell R. W. Three cysteine residues in the alpha chain of rat haemoglobin (albino Rattus norvegicus): 13 (A 11), 104 (G 11) and 111 (G 18). Biochim Biophys Acta. 1974 Oct 9;365(2):328–334. doi: 10.1016/0005-2795(74)90005-1. [DOI] [PubMed] [Google Scholar]
  3. Eriksson C. J., Sippel H. W., Forsander O. A. The occurrence of acetaldehyde binding in rat blood but not in human blood. FEBS Lett. 1977 Mar 15;75(1):205–208. doi: 10.1016/0014-5793(77)80087-2. [DOI] [PubMed] [Google Scholar]
  4. Garrick L. M., Sharma V. S., McDonald M. J., Ranney H. M. Rat haemoglobin heterogeneity. Two structurally distinct alpha chains and functional behaviour of selected components. Biochem J. 1975 Jul;149(1):245–258. doi: 10.1042/bj1490245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Garrick L. M., Sloan R. L., Ryan T. W., Klonowski T. J., Garrick M. D. Primary structure of the major beta-chain of rat haemoglobins. Biochem J. 1978 Jul 1;173(1):321–330. doi: 10.1042/bj1730321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Honegger A., Hughes G. J., Wilson K. J. Chemical modification of peptides by hydrazine. Biochem J. 1981 Oct 1;199(1):53–59. doi: 10.1042/bj1990053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hughes G. J., Winterhalter K. H., Lutz H., Wilson K. J. Microsequence analysis: III. Automatic solid-phage sequencing using DABITC. FEBS Lett. 1979 Dec 1;108(1):92–97. doi: 10.1016/0014-5793(79)81185-0. [DOI] [PubMed] [Google Scholar]
  8. Hughes G. J., Winterhalter K. H., Wilson K. J. Microsequence analysis: I. Peptide isolation using high-performance liquid chromatography. FEBS Lett. 1979 Dec 1;108(1):81–86. doi: 10.1016/0014-5793(79)81183-7. [DOI] [PubMed] [Google Scholar]
  9. Kosower N. S., Kosower E. M., Koppel R. L. Sensitivity of hemoglobin thiol groups within red blood cells of rat during oxidation of glutathione. Eur J Biochem. 1977 Aug 1;77(3):529–534. doi: 10.1111/j.1432-1033.1977.tb11695.x. [DOI] [PubMed] [Google Scholar]
  10. Perutz M. F., Muirhead H., Cox J. M., Goaman L. C. Three-dimensional Fourier synthesis of horse oxyhaemoglobin at 2.8 A resolution: the atomic model. Nature. 1968 Jul 13;219(5150):131–139. doi: 10.1038/219131a0. [DOI] [PubMed] [Google Scholar]
  11. Wilson K. J., Honegger A., Hughes G. J. Comparison of buffers and detection systems for high-pressure liquid chromatography of peptide mixtures. Biochem J. 1981 Oct 1;199(1):43–51. doi: 10.1042/bj1990043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Wilson K. J., Honegger A., Stötzel R. P., Hughes G. J. The behaviour of peptides on reverse-phase supports during high-pressure liquid chromatography. Biochem J. 1981 Oct 1;199(1):31–41. doi: 10.1042/bj1990031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wilson K. J., Rodger K., Hughes G. J. Microsequence analyses: II. DABTH-amino acid identification by high-performance liquid and thin-layer chromatography. FEBS Lett. 1979 Dec 1;108(1):87–91. doi: 10.1016/0014-5793(79)81184-9. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES