Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1986 Mar 15;234(3):649–652. doi: 10.1042/bj2340649

Identification and partial characterization of the human erythrocyte membrane component(s) that express the antigens of the LW blood-group system.

G Mallinson, P G Martin, D J Anstee, M J Tanner, A H Merry, D Tills, H H Sonneborn
PMCID: PMC1146620  PMID: 2424433

Abstract

Rhnull human erythrocytes lack the antigens of the Rhesus blood-group system, have an abnormal shape, have an increased osmotic fragility, and are associated with mild chronic haemolytic anaemia. Rhnull erythrocytes also lack all antigens of the LW blood-group system, but the functional significance of this deficiency is unknown. We have identified, by immunoblotting with two mouse monoclonal antibodies (BS46 and BS56), the LW-active component(s) in normal human erythrocytes as a broad band of Mr 37 000-47 000 on SDS/polyacrylamide-gel electrophoresis. Treatment of intact human erythrocytes with endoglycosidase F preparation destroyed the epitopes recognized by antibodies BS46 and BS56, suggesting that one or more N-glycosidically linked oligosaccharides are required for the formation of the LW antigens. Estimation of the number of LW antigen sites per erythrocyte by using radioiodinated purified antibody BS46 gave average values of 4400 molecules/cell for Rh(D)-positive adult erythrocytes and 2835 molecules/cell for Rh(D)-negative adult erythrocytes. Like the Rh(D) polypeptide, the LW polypeptide(s) is (are) associated with the cytoskeleton of normal erythrocytes. These results suggest the possibility that the absence of the LW polypeptide may also contribute to the functional and/or morphological abnormalities of Rhnull erythrocytes.

Full text

PDF
649

Images in this article

651Fig. 1.
on p.651

Selected References

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

  1. Anstee D. J., Mawby W. J., Tanner M. J. Abnormal blood-group-Ss-active sialoglycoproteins in the membrane of Miltenberger class III, IV and V human erythrocytes. Biochem J. 1979 Nov 1;183(2):193–203. doi: 10.1042/bj1830193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anstee D. J., Parsons S. F., Ridgwell K., Tanner M. J., Merry A. H., Thomson E. E., Judson P. A., Johnson P., Bates S., Fraser I. D. Two individuals with elliptocytic red cells apparently lack three minor erythrocyte membrane sialoglycoproteins. Biochem J. 1984 Mar 1;218(2):615–619. doi: 10.1042/bj2180615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Batteiger B., Newhall W. J., 5th, Jones R. B. The use of Tween 20 as a blocking agent in the immunological detection of proteins transferred to nitrocellulose membranes. J Immunol Methods. 1982 Dec 30;55(3):297–307. doi: 10.1016/0022-1759(82)90089-8. [DOI] [PubMed] [Google Scholar]
  4. Doyle A., Jones T. J., Bidwell J. L., Bradley B. A. In vitro development of human monoclonal antibody-secreting plasmacytomas. Hum Immunol. 1985 Jul;13(3):199–209. doi: 10.1016/0198-8859(85)90012-6. [DOI] [PubMed] [Google Scholar]
  5. Elder J. H., Alexander S. endo-beta-N-acetylglucosaminidase F: endoglycosidase from Flavobacterium meningosepticum that cleaves both high-mannose and complex glycoproteins. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4540–4544. doi: 10.1073/pnas.79.15.4540. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fukuda M., Fukuda M. N. Changes in cell surface glycoproteins and carbohydrate structures during the development and differentiation of human erythroid cells. J Supramol Struct Cell Biochem. 1981;17(4):313–324. doi: 10.1002/jsscb.380170403. [DOI] [PubMed] [Google Scholar]
  7. Gahmberg C. G., Karhi K. K. Association of Rho(D) polypeptides with the membrane skeleton in Rho(D)-positive human red cells. J Immunol. 1984 Jul;133(1):334–337. [PubMed] [Google Scholar]
  8. Gahmberg C. G. Molecular characterization of the human red cell Rho(D) antigen. EMBO J. 1983;2(2):223–227. doi: 10.1002/j.1460-2075.1983.tb01409.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gahmberg C. G. Molecular identification of the human Rho (D) antigen. FEBS Lett. 1982 Apr 5;140(1):93–97. doi: 10.1016/0014-5793(82)80528-0. [DOI] [PubMed] [Google Scholar]
  10. Kitamikado M., Ito M., Li Y. T. Isolation and characterization of a keratan sulfate-degrading endo-beta-galactosidase from Flavobacterium keratolyticus. J Biol Chem. 1981 Apr 25;256(8):3906–3909. [PubMed] [Google Scholar]
  11. Konigshaus G. J., Holland T. I. The effect of dithiothreitol on the LW antigen. Transfusion. 1984 Nov-Dec;24(6):536–537. doi: 10.1046/j.1537-2995.1984.24685066821.x. [DOI] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Lomas C. G., Tippett P. Use of enzymes in distinguishing anti-LWa and anti-LWab from anti-D. Med Lab Sci. 1985 Jan;42(1):88–89. [PubMed] [Google Scholar]
  14. Merry A. H., Hodson C., Thomson E., Mallinson G., Anstee D. J. The use of monoclonal antibodies to quantify the levels of sialoglycoproteins alpha and delta and variant sialoglycoproteins in human erythrocyte membranes. Biochem J. 1986 Jan 1;233(1):93–98. doi: 10.1042/bj2330093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Moore S., Woodrow C. F., McClelland D. B. Isolation of membrane components associated with human red cell antigens Rh(D), (c), (E) and Fy. Nature. 1982 Feb 11;295(5849):529–531. doi: 10.1038/295529a0. [DOI] [PubMed] [Google Scholar]
  16. Plummer T. H., Jr, Elder J. H., Alexander S., Phelan A. W., Tarentino A. L. Demonstration of peptide:N-glycosidase F activity in endo-beta-N-acetylglucosaminidase F preparations. J Biol Chem. 1984 Sep 10;259(17):10700–10704. [PubMed] [Google Scholar]
  17. Ridgwell K., Roberts S. J., Tanner M. J., Anstee D. J. Absence of two membrane proteins containing extracellular thiol groups in Rhnull human erythrocytes. Biochem J. 1983 Jul 1;213(1):267–269. doi: 10.1042/bj2130267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ridgwell K., Tanner M. J., Anstee D. J. The Rhesus (D) polypeptide is linked to the human erythrocyte cytoskeleton. FEBS Lett. 1984 Aug 20;174(1):7–10. doi: 10.1016/0014-5793(84)81066-2. [DOI] [PubMed] [Google Scholar]
  19. Sistonen P., Green C. A., Lomas C. G., Tippett P. Genetic polymorphism of the LW blood group system. Ann Hum Genet. 1983 Oct;47(Pt 4):277–284. doi: 10.1111/j.1469-1809.1983.tb00997.x. [DOI] [PubMed] [Google Scholar]
  20. Tanner M. J., Anstee D. J., Mawby W. J. A new human erythrocyte variant (Ph) containing an abnormal membrane sialoglycoprotein. Biochem J. 1980 May 1;187(2):493–500. doi: 10.1042/bj1870493. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES