Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1971 Dec;125(4):1109–1117. doi: 10.1042/bj1251109

The isolation and functional identification of a protein from the human erythrocyte `ghost'

M J A Tanner 1, W R Gray 1,*
PMCID: PMC1178275  PMID: 5004149

Abstract

A protein, initially identified as a band on polyacrylamide-gel electrophoresis of erythrocyte `ghosts', was isolated by selective extraction of `ghosts' with EDTA solutions. The molecular weight of the polypeptide chain was estimated as 33000 and it represents approx. 5% of the membrane protein. The N-terminal sequence of the protein was established. Comparison with known protein sequences suggested that the protein might be the erythrocyte d-glyceraldehyde 3-phosphate dehydrogenase. This identification was confirmed by direct enzyme assay. It is suggested that this enzyme, which is strongly retained by erythrocyte `ghosts' on haemolysis of erythrocytes, is unlikely to be an integral part of the structure of the erythrocyte membrane.

Full text

PDF

Images in this article

Selected References

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

  1. Bakerman S., Wasemiller G. Studies on structural units of human erythrocyte membrane. I. Separation, isolation, and partial characterization. Biochemistry. 1967 Apr;6(4):1100–1113. doi: 10.1021/bi00856a020. [DOI] [PubMed] [Google Scholar]
  2. DODGE J. T., MITCHELL C., HANAHAN D. J. The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch Biochem Biophys. 1963 Jan;100:119–130. doi: 10.1016/0003-9861(63)90042-0. [DOI] [PubMed] [Google Scholar]
  3. Edman P., Begg G. A protein sequenator. Eur J Biochem. 1967 Mar;1(1):80–91. doi: 10.1007/978-3-662-25813-2_14. [DOI] [PubMed] [Google Scholar]
  4. Green D. E., Murer E., Hultin H. O., Richardson S. H., Salmon B., Brierley G. P., Baum H. Association of integrated metabolic pathways with membranes. I. Glycolytic enzymes of the red blood corpuscle and yeast. Arch Biochem Biophys. 1965 Dec;112(3):635–647. doi: 10.1016/0003-9861(65)90107-4. [DOI] [PubMed] [Google Scholar]
  5. HASHIMOTO T., YOSHIKAWA H. HUMAN ERYTHROCYTE ADENOSINE TRIPHOSPHATE. D-3-PHOSPHOGLYCERATE 1-PHOSPHOTRANSFERASE. J Biochem. 1964 Sep;56:279–284. doi: 10.1093/oxfordjournals.jbchem.a127990. [DOI] [PubMed] [Google Scholar]
  6. Harris J. I., Perham R. N. Glyceraldehyde 3-phosphate dehydrogenase from pig muscle. Nature. 1968 Sep 7;219(5158):1025–1028. doi: 10.1038/2191025a0. [DOI] [PubMed] [Google Scholar]
  7. KATZ A. M., DREYER W. J., ANFINSEN C. B. Peptide separation by two-dimensional chromatography and electrophoresis. J Biol Chem. 1959 Nov;234:2897–2900. [PubMed] [Google Scholar]
  8. Lewis W. H., Harris H. Human red cell peptidases. Nature. 1967 Jul 22;215(5099):351–355. doi: 10.1038/215351a0. [DOI] [PubMed] [Google Scholar]
  9. MORRISON W. L., NEURATH H. Proteolytic enzymes of the formed elements of human blood. I. Erythrocytes. J Biol Chem. 1953 Jan;200(1):39–51. [PubMed] [Google Scholar]
  10. Mitchell C. D., Mitchell W. B., Hanahan D. J. Enzyme and hemoglobin retention in human erythrocyte stroma. Biochim Biophys Acta. 1965 Jul 8;104(2):348–358. doi: 10.1016/0304-4165(65)90340-5. [DOI] [PubMed] [Google Scholar]
  11. Moore G. L., Kocholaty W. F., Cooper D. A., Gray J. L., Robinson S. L. A proteinase from human erythrocyte membranes. Biochim Biophys Acta. 1970 Jul 15;212(1):126–133. doi: 10.1016/0005-2744(70)90185-3. [DOI] [PubMed] [Google Scholar]
  12. Neville D. M., Jr Fractionation of cell membrane protein by disc electrophoresis. Biochim Biophys Acta. 1967 Jan 18;133(1):168–170. doi: 10.1016/0005-2795(67)90051-7. [DOI] [PubMed] [Google Scholar]
  13. Rosenberg S. A., Guidotti G. Fractionation of the protein components of human erythrocyte membranes. J Biol Chem. 1969 Oct 10;244(19):5118–5124. [PubMed] [Google Scholar]
  14. Schneiderman L. J., Junga I. G. Isolation and partial characterization of structural protein derived from human red cell membranes. Biochemistry. 1968 Jun;7(6):2281–2286. doi: 10.1021/bi00846a034. [DOI] [PubMed] [Google Scholar]
  15. Shapiro A. L., Viñuela E., Maizel J. V., Jr Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun. 1967 Sep 7;28(5):815–820. doi: 10.1016/0006-291x(67)90391-9. [DOI] [PubMed] [Google Scholar]
  16. Wolny M., Wolny E., Baranowski T. Preparation and some properties of crystalline D-glyceraldehyde-3-phosphate dehydrogenase from human erythrocytes. Bull Acad Pol Sci Biol. 1968;16(1):13–16. [PubMed] [Google Scholar]
  17. Zacharius R. M., Zell T. E., Morrison J. H., Woodlock J. J. Glycoprotein staining following electrophoresis on acrylamide gels. Anal Biochem. 1969 Jul;30(1):148–152. doi: 10.1016/0003-2697(69)90383-2. [DOI] [PubMed] [Google Scholar]

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

RESOURCES