Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1986 Jan 1;233(1):93–98. doi: 10.1042/bj2330093

The use of monoclonal antibodies to quantify the levels of sialoglycoproteins alpha and delta and variant sialoglycoproteins in human erythrocyte membranes.

A H Merry, C Hodson, E Thomson, G Mallinson, D J Anstee
PMCID: PMC1152989  PMID: 2420325

Abstract

By using radioiodinated monoclonal antibodies we have estimated that there are about 600 000 copies of sialoglycoprotein alpha (synonym glycophorin A) and 80 000 copies of sialoglycoprotein delta (synonym glycophorin B) per normal human erythrocyte. Erythrocytes expressing the product of only one alpha gene contain about 300 000 copies of alpha/cell. Two erythrocyte types containing alpha-delta hybrid molecules were studied. Those with heterozygous expression of the (alpha-delta)Mi.V gene contain about 100 000 alpha-delta copies per cell, whereas those with heterozygous expression of the En(UK) gene contain about 80 000 alpha-delta copies/cell. Erythrocyte types containing delta-alpha hybrid molecules were also studied. About 200 000 copies of (delta-alpha)Dantu were measured in cells with heterozygous expression of the (delta-alpha)Dantu gene (donor M.P.), whereas about 315 000 copies of the putative (delta-alpha)Dantu hybrid were found on the erythrocytes of donor J.O. [which also have heterozygous expression of the putative (delta-alpha)Dantu gene]. The erythrocytes of donor M.P. have normal levels of alpha, whereas those of donor J.O. have only about half-normal levels. It is proposed that the hybrid sialoglycoprotein of donor J.O. is of alpha-delta-alpha composition [(alpha-delta-alpha)Dantu] rather than delta-alpha and results from a double cross-over analogous to that which gives rise to haemoglobin Parchman.

Full text

PDF
93

Images in this article

Selected References

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

  1. Adams J. G., 3rd, Morrison W. T., Steinberg M. H. Hemoglobin Parchman: double crossover within a single human gene. Science. 1982 Oct 15;218(4569):291–293. doi: 10.1126/science.7123235. [DOI] [PubMed] [Google Scholar]
  2. Anstee D. J., Edwards P. A. Monoclonal antibodies to human erythrocytes. Eur J Immunol. 1982 Mar;12(3):228–232. doi: 10.1002/eji.1830120311. [DOI] [PubMed] [Google Scholar]
  3. Anstee D. J., Mawby W. J., Parsons S. F., Tanner M. J., Giles C. M. A novel hybrid sialoglycoprotein in Sta positive human erythrocytes. J Immunogenet. 1982 Feb;9(1):51–55. doi: 10.1111/j.1744-313x.1982.tb00782.x. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. 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]
  7. Contreras M., Green C., Humphreys J., Tippett P., Daniels G., Teesdale P., Armitage S., Lubenko A. Serology and genetics of an MNSs-associated antigen Dantu. Vox Sang. 1984;46(6):377–386. doi: 10.1111/j.1423-0410.1984.tb00102.x. [DOI] [PubMed] [Google Scholar]
  8. Dahr W., Uhlenbruck G., Leikola J., Wagstaff W. Studies on the membrane glycoprotein defect of En(a-) erythrocytes. III. N-terminal amino acids of sialoglycoproteins from normal and En(a-) red cells. J Immunogenet. 1978 Apr;5(2):117–127. doi: 10.1111/j.1744-313x.1978.tb00636.x. [DOI] [PubMed] [Google Scholar]
  9. Edwards P. A. Monoclonal antibodies that bind to the human erythrocyte-membrane glycoproteins glycophorin A and Band 3 [proceedings]. Biochem Soc Trans. 1980 Jun;8(3):334–335. doi: 10.1042/bst0080334. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Gahmberg C. G., Jokinen M., Andersson L. C. Expression of the major red cell sialoglycoprotein, glycophorin A, in the human leukemic cell line K562. J Biol Chem. 1979 Aug 10;254(15):7442–7448. [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. Mawby W. J., Anstee D. J., Tanner M. J. Immunochemical evidence for hybrid sialoglycoproteins of human erythrocytes. Nature. 1981 May 14;291(5811):161–162. doi: 10.1038/291161a0. [DOI] [PubMed] [Google Scholar]
  14. Merry A. H., Thomson E. E., Anstee D. J., Stratton F. The quantification of erythrocyte antigen sites with monoclonal antibodies. Immunology. 1984 Apr;51(4):793–800. [PMC free article] [PubMed] [Google Scholar]
  15. Modiano G., Pepe G. The quantitative expression of delta and beta human globin genes is controlled by both 5' and 3' untranslated regions. Mol Biol Med. 1983 Jul;1(1):157–159. [PubMed] [Google Scholar]
  16. Perry R. P. What controls the transcription of immunoglobulin genes? Nature. 1984 Jul 5;310(5972):14–15. doi: 10.1038/310014a0. [DOI] [PubMed] [Google Scholar]
  17. Ridgwell K., Tanner M. J., Anstee D. J. The Wrb antigen in Sta-positive and Dantu-positive human erythrocytes. J Immunogenet. 1984 Oct-Dec;11(5-6):365–370. doi: 10.1111/j.1744-313x.1984.tb00822.x. [DOI] [PubMed] [Google Scholar]
  18. Ridgwell K., Tanner M. J., Anstee D. J. The Wrb antigen, a receptor for Plasmodium falciparum malaria, is located on a helical region of the major membrane sialoglycoprotein of human red blood cells. Biochem J. 1983 Jan 1;209(1):273–276. doi: 10.1042/bj2090273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Steck T. L. The organization of proteins in the human red blood cell membrane. A review. J Cell Biol. 1974 Jul;62(1):1–19. doi: 10.1083/jcb.62.1.1. [DOI] [PMC free article] [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]
  21. Vengelen-Tyler V., Anstee D. J., Issitt P. D., Pavone B. G., Ferguson S. J., Mawby W. J., Tanner M. J., Blajchman M. A., Lorque P. Studies on the blood of an MiV homozygote. Transfusion. 1981 Jan-Feb;21(1):1–14. [PubMed] [Google Scholar]

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

RESOURCES