Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1972 Sep;129(2):333–347. doi: 10.1042/bj1290333

Separation and some properties of the major proteins of the human erythrocyte membrane

M J A Tanner 1, D H Boxer 1
PMCID: PMC1174083  PMID: 4643321

Abstract

A fractionation procedure is described which allows the isolation of three major human erythrocyte membrane proteins. Their isolation involves three sequential extraction procedures followed by gel filtration in 1% sodium dodecyl sulphate and preparative gel electrophoresis. All three proteins can be isolated from a single preparation. One of the proteins is the erythrocyte sialoglycoprotein, for which no C- or N-terminal residues were found. The other two proteins, which have not previously been isolated, have subunit molecular weights of 74000 and 93000 and contain 9 and 7% carbohydrate respectively. These glycoproteins have blocked N-terminal residues and show similarities in their chemical properties. Preparations derived from blood-group O erythrocytes contain no N-acetylgalactosamine, but similar preparations from blood-group A erythrocytes do contain this sugar. These three proteins cannot easily be solubilized by gentle aqueous procedures and represent about half of the erythrocyte `ghost' protein. They carry a large proportion of the cell-surface carbohydrate.

Full text

PDF
335

Images in this article

Selected References

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

  1. BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
  2. Bellhorn M. B., Blumenfeld O. O., Gallop P. M. Acetylcholinesterase of the human erythrocyte membrane. Biochem Biophys Res Commun. 1970 Apr 24;39(2):267–273. doi: 10.1016/0006-291x(70)90788-6. [DOI] [PubMed] [Google Scholar]
  3. Bender W. W., Garan H., Berg H. C. Proteins of the human erythrocyte membrane as modified by pronase. J Mol Biol. 1971 Jun 28;58(3):783–797. doi: 10.1016/0022-2836(71)90040-4. [DOI] [PubMed] [Google Scholar]
  4. Berg H. C. Sulfanilic acid diazonium salt: a label for the outside of the human erythrocyte membrane. Biochim Biophys Acta. 1969 Jun 3;183(1):65–78. doi: 10.1016/0005-2736(69)90130-8. [DOI] [PubMed] [Google Scholar]
  5. Bhatti T., Chambers R. E., Clamp J. R. The gas chromatographic properties of biologically important N-acetylglucosamine derivatives, monosaccharides, disaccharides, trisaccharides, tetrasaccharides and pentasaccharides. Biochim Biophys Acta. 1970 Nov 24;222(2):339–347. doi: 10.1016/0304-4165(70)90122-4. [DOI] [PubMed] [Google Scholar]
  6. Blumenfeld O. O. The proteins of the erythrocyte membrane obtained by solubilization with aqueous pyridine solution. Biochem Biophys Res Commun. 1968 Jan 25;30(2):200–205. doi: 10.1016/0006-291x(68)90471-3. [DOI] [PubMed] [Google Scholar]
  7. Bretscher M. S. A major protein which spans the human erythrocyte membrane. J Mol Biol. 1971 Jul 28;59(2):351–357. doi: 10.1016/0022-2836(71)90055-6. [DOI] [PubMed] [Google Scholar]
  8. Bretscher M. S. Human erythrocyte membranes: specific labelling of surface proteins. J Mol Biol. 1971 Jun 28;58(3):775–781. doi: 10.1016/0022-2836(71)90039-8. [DOI] [PubMed] [Google Scholar]
  9. Bretscher M. S. Major human erythrocyte glycoprotein spans the cell membrane. Nat New Biol. 1971 Jun 23;231(25):229–232. doi: 10.1038/newbio231229a0. [DOI] [PubMed] [Google Scholar]
  10. Brownstone A. D. A versatile system for preparative electrophoresis in acrylamide gel. Anal Biochem. 1969 Jan;27(1):25–46. doi: 10.1016/0003-2697(69)90216-4. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Dunham P. B., Hoffman J. F. Partial purification of the ouabain-binding component and of Na,K-ATPase from human red cell membranes. Proc Natl Acad Sci U S A. 1970 Jul;66(3):936–943. doi: 10.1073/pnas.66.3.936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  14. GUIDOTTI G. The action of carboxypeptidases A and B on the separated alpha and beta chains of normal adult human hemoglobin. Biochim Biophys Acta. 1960 Jul 29;42:177–179. doi: 10.1016/0006-3002(60)90774-5. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Green D. E., Perdue J. F. Membranes as expressions of repeating units. Proc Natl Acad Sci U S A. 1966 May;55(5):1295–1302. doi: 10.1073/pnas.55.5.1295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lenard J. Protein and glycolipid components of human erythrocyte membranes. Biochemistry. 1970 Mar 3;9(5):1129–1132. doi: 10.1021/bi00807a012. [DOI] [PubMed] [Google Scholar]
  18. MORAWIECKI A. DISSOCIATION OF M- AND N-GROUP MUCOPROTEINS INTO SUBUNITS IN DETERGENT SOLUTIONS. Biochim Biophys Acta. 1964 Nov 1;83:339–347. doi: 10.1016/0926-6526(64)90012-6. [DOI] [PubMed] [Google Scholar]
  19. Marchesi S. L., Steers E., Marchesi V. T., Tillack T. W. Physical and chemical properties of a protein isolated from red cell membranes. Biochemistry. 1970 Jan 6;9(1):50–57. doi: 10.1021/bi00803a007. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Phillips D. R., Morrison M. Exposed protein on the intact human erythrocyte. Biochemistry. 1971 May 11;10(10):1766–1771. doi: 10.1021/bi00786a006. [DOI] [PubMed] [Google Scholar]
  22. Phillips D. R., Morrison M. Exterior proteins on the human erythrocyte membrane. Biochem Biophys Res Commun. 1971 Nov;45(4):1103–1108. doi: 10.1016/0006-291x(71)90451-7. [DOI] [PubMed] [Google Scholar]
  23. Segrest J. P., Jackson R. L., Andrews E. P., Marchesi V. T. Human erythrocyte membrane glycoprotein: a re-evaluation of the molecular weight as determined by SDS polyacrylamide gel electrophoresis. Biochem Biophys Res Commun. 1971 Jul 16;44(2):390–395. doi: 10.1016/0006-291x(71)90612-7. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Springer G. F., Nagai Y., Tegtmeyer H. Isolation and properties of human blood-group NN and meconium-Vg antigens. Biochemistry. 1966 Oct;5(10):3254–3272. doi: 10.1021/bi00874a028. [DOI] [PubMed] [Google Scholar]
  26. Steck T. L., Fairbanks G., Wallach D. F. Disposition of the major proteins in the isolated erythrocyte membrane. Proteolytic dissection. Biochemistry. 1971 Jun 22;10(13):2617–2624. doi: 10.1021/bi00789a031. [DOI] [PubMed] [Google Scholar]
  27. Tanner M. J., Gray W. R. The isolation and functional identification of a protein from the human erythrocyte 'ghost'. Biochem J. 1971 Dec;125(4):1109–1117. doi: 10.1042/bj1251109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tillack T. W., Marchesi V. T. Demonstration of the outer surface of freeze-etched red blood cell membranes. J Cell Biol. 1970 Jun;45(3):649–653. doi: 10.1083/jcb.45.3.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. WARREN L. The thiobarbituric acid assay of sialic acids. J Biol Chem. 1959 Aug;234(8):1971–1975. [PubMed] [Google Scholar]
  30. Winzler R. J. Carbohydrates in cell surfaces. Int Rev Cytol. 1970;29:77–125. doi: 10.1016/s0074-7696(08)60033-9. [DOI] [PubMed] [Google Scholar]
  31. ZLATKIS A., ZAK B., BOYLE A. J. A new method for the direct determination of serum cholesterol. J Lab Clin Med. 1953 Mar;41(3):486–492. [PubMed] [Google Scholar]
  32. 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]
  33. Zvilichovsky B., Gallop P. M., Blumenfeld O. O. Isolation of a glycoprotein--glycolipid fraction from human erythrocyte membranes. Biochem Biophys Res Commun. 1971 Sep;44(5):1234–1243. doi: 10.1016/s0006-291x(71)80218-8. [DOI] [PubMed] [Google Scholar]

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

RESOURCES