Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1987 May;6(5):1259–1264. doi: 10.1002/j.1460-2075.1987.tb02362.x

Lymphocyte specific heterogeneity in the rat leucocyte common antigen (T200) is due to differences in polypeptide sequences near the NH2-terminus.

A N Barclay, D I Jackson, A C Willis, A F Williams
PMCID: PMC553927  PMID: 2440674

Abstract

The leucocyte-common antigen (L-CA, T200 or CD45) consists of a family of heavily glycosylated glycoproteins of apparent Mr 180,000-240,000 which are restricted to lymphoid and myeloid cells. Forms of L-CA which differ in their apparent Mr, antigenicity and glycosylation are expressed on different lymphocyte types. One specific antigenic determinant called MRC OX-22 is of particular interest because it distinguishes two sets of T helper cells that have different functions. From the sequence of different L-CA cDNA clones we now conclude that there is sequence heterogeneity such that at least four forms of L-CA exist with sequences in the range 1118-1250 amino acids. All the sequence variation occurs at a point starting 6 residues from the NH2-terminus and the last 1112 residues of all forms are identical. Two of the variants can be directly related to the antigenic variation because they include sequence that was determined for a peptide that carries the MRC OX-22 determinant. Analysis of glycopeptides from thymocyte L-CA identified only one non-glycosylated position out of 14 possible N-glycosylation sites and established that all O-glycosylation was within the first 32 amino acids. The extra protein sequence in the longer forms was also suggestive of extensive O-glycosylation.

Full text

PDF
1259

Images in this article

Selected References

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

  1. Arthur R. P., Mason D. T cells that help B cell responses to soluble antigen are distinguishable from those producing interleukin 2 on mitogenic or allogeneic stimulation. J Exp Med. 1986 Apr 1;163(4):774–786. doi: 10.1084/jem.163.4.774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bause E., Hettkamp H. Primary structural requirements for N-glycosylation of peptides in rat liver. FEBS Lett. 1979 Dec 15;108(2):341–344. doi: 10.1016/0014-5793(79)80559-1. [DOI] [PubMed] [Google Scholar]
  3. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bourguignon L. Y., Suchard S. J., Nagpal M. L., Glenney J. R., Jr A T-lymphoma transmembrane glycoprotein (gp180) is linked to the cytoskeletal protein, fodrin. J Cell Biol. 1985 Aug;101(2):477–487. doi: 10.1083/jcb.101.2.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Breitbart R. E., Nguyen H. T., Medford R. M., Destree A. T., Mahdavi V., Nadal-Ginard B. Intricate combinatorial patterns of exon splicing generate multiple regulated troponin T isoforms from a single gene. Cell. 1985 May;41(1):67–82. doi: 10.1016/0092-8674(85)90062-5. [DOI] [PubMed] [Google Scholar]
  6. Brown W. R., Barclay A. N., Sunderland C. A., Williams A. F. Identification of a glycophorin-like molecule at the cell surface of rat thymocytes. Nature. 1981 Feb 5;289(5797):456–460. doi: 10.1038/289456a0. [DOI] [PubMed] [Google Scholar]
  7. Brown W. R., Williams A. F. Lymphocyte cell surface glycoproteins which bind to soybean and peanut lectins. Immunology. 1982 Aug;46(4):713–726. [PMC free article] [PubMed] [Google Scholar]
  8. Campbell D. G., Gagnon J., Reid K. B., Williams A. F. Rat brain Thy-1 glycoprotein. The amino acid sequence, disulphide bonds and an unusual hydrophobic region. Biochem J. 1981 Apr 1;195(1):15–30. doi: 10.1042/bj1950015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Childs R. A., Dalchau R., Scudder P., Hounsell E. F., Fabre J. W., Feizi T. Evidence for the occurrence of O-glycosidically linked oligosaccharides of poly-N-acetyllactosamine type on the human leucocyte common antigen. Biochem Biophys Res Commun. 1983 Jan 27;110(2):424–431. doi: 10.1016/0006-291x(83)91166-x. [DOI] [PubMed] [Google Scholar]
  10. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  11. Coffman R. L., Weissman I. L. B220: a B cell-specific member of th T200 glycoprotein family. Nature. 1981 Feb 19;289(5799):681–683. doi: 10.1038/289681a0. [DOI] [PubMed] [Google Scholar]
  12. Dalchau R., Fabre J. W. Identification with a monoclonal antibody of a predominantly B lymphocyte-specific determinant of the human leukocyte common antigen. Evidence for structural and possible functional diversity of the human leukocyte common molecule. J Exp Med. 1981 Apr 1;153(4):753–765. doi: 10.1084/jem.153.4.753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dalchau R., Kirkley J., Fabre J. W. Monoclonal antibody to a human leukocyte-specific membrane glycoprotein probably homologous to the leukocyte-common (L-C) antigen of the rat. Eur J Immunol. 1980 Oct;10(10):737–744. doi: 10.1002/eji.1830101003. [DOI] [PubMed] [Google Scholar]
  14. Fabre J. W., Williams A. F. Quantitative serological analysis of a rabbit anti-rat lymphocyte serum and preliminary biochemical characterisation of the major antigen recognised. Transplantation. 1977 Apr;23(4):349–359. doi: 10.1097/00007890-197704000-00009. [DOI] [PubMed] [Google Scholar]
  15. Frangione B., Rosenwasser E., Prelli F., Franklin E. C. Primary structure of human gamma 3 immunoglobulin deletion mutant: gamma 3 heavy-chain disease protein Wis. Biochemistry. 1980 Sep 2;19(18):4304–4308. doi: 10.1021/bi00559a024. [DOI] [PubMed] [Google Scholar]
  16. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  17. Johnson P., Gagnon J., Barclay A. N., Williams A. F. Purification, chain separation and sequence of the MRC OX-8 antigen, a marker of rat cytotoxic T lymphocytes. EMBO J. 1985 Oct;4(10):2539–2545. doi: 10.1002/j.1460-2075.1985.tb03968.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kornblihtt A. R., Umezawa K., Vibe-Pedersen K., Baralle F. E. Primary structure of human fibronectin: differential splicing may generate at least 10 polypeptides from a single gene. EMBO J. 1985 Jul;4(7):1755–1759. doi: 10.1002/j.1460-2075.1985.tb03847.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lefrancois L., Puddington L., Machamer C. E., Bevan M. J. Acquisition of cytotoxic T lymphocyte-specific carbohydrate differentiation antigens. J Exp Med. 1985 Oct 1;162(4):1275–1293. doi: 10.1084/jem.162.4.1275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lefrancois L., Thomas M. L., Bevan M. J., Trowbridge I. S. Different classes of T lymphocytes have different mRNAs for the leukocyte-common antigen, T200. J Exp Med. 1986 May 1;163(5):1337–1342. doi: 10.1084/jem.163.5.1337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. McMullen B. A., Fujikawa K. Amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor). J Biol Chem. 1985 May 10;260(9):5328–5341. [PubMed] [Google Scholar]
  22. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  23. Murray B. A., Owens G. C., Prediger E. A., Crossin K. L., Cunningham B. A., Edelman G. M. Cell surface modulation of the neural cell adhesion molecule resulting from alternative mRNA splicing in a tissue-specific developmental sequence. J Cell Biol. 1986 Oct;103(4):1431–1439. doi: 10.1083/jcb.103.4.1431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nawa H., Kotani H., Nakanishi S. Tissue-specific generation of two preprotachykinin mRNAs from one gene by alternative RNA splicing. Nature. 1984 Dec 20;312(5996):729–734. doi: 10.1038/312729a0. [DOI] [PubMed] [Google Scholar]
  25. Newton M. R., Wood K. J., Fabre J. W. A monoclonal alloantibody detecting a polymorphism of the rat leucocyte common (LC) antigen. J Immunogenet. 1986 Feb;13(1):41–50. doi: 10.1111/j.1744-313x.1986.tb01081.x. [DOI] [PubMed] [Google Scholar]
  26. Podell D. N., Abraham G. N. A technique for the removal of pyroglutamic acid from the amino terminus of proteins using calf liver pyroglutamate amino peptidase. Biochem Biophys Res Commun. 1978 Mar 15;81(1):176–185. doi: 10.1016/0006-291x(78)91646-7. [DOI] [PubMed] [Google Scholar]
  27. Ralph S. J., Thomas M. L., Morton C. C., Trowbridge I. S. Structural variants of human T200 glycoprotein (leukocyte-common antigen). EMBO J. 1987 May;6(5):1251–1257. doi: 10.1002/j.1460-2075.1987.tb02361.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Saga Y., Tung J. S., Shen F. W., Boyse E. A. Sequences of Ly-5 cDNA: isoform-related diversity of Ly-5 mRNA. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6940–6944. doi: 10.1073/pnas.83.18.6940. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Shackelford D. A., Trowbridge I. S. Identification of lymphocyte integral membrane proteins as substrates for protein kinase C. Phosphorylation of the interleukin-2 receptor, class I HLA antigens, and T200 glycoprotein. J Biol Chem. 1986 Jun 25;261(18):8334–8341. [PubMed] [Google Scholar]
  30. Spickett G. P., Brandon M. R., Mason D. W., Williams A. F., Woollett G. R. MRC OX-22, a monoclonal antibody that labels a new subset of T lymphocytes and reacts with the high molecular weight form of the leukocyte-common antigen. J Exp Med. 1983 Sep 1;158(3):795–810. doi: 10.1084/jem.158.3.795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Staden R. The current status and portability of our sequence handling software. Nucleic Acids Res. 1986 Jan 10;14(1):217–231. doi: 10.1093/nar/14.1.217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Standring R., McMaster W. R., Sunderland C. A., Williams A. F. The predominant heavily glycosylated glycoproteins at the surface of rat lymphoid cells are differentiation antigens. Eur J Immunol. 1978 Dec;8(12):832–839. doi: 10.1002/eji.1830081203. [DOI] [PubMed] [Google Scholar]
  33. Thomas M. L., Barclay A. N., Gagnon J., Williams A. F. Evidence from cDNA clones that the rat leukocyte-common antigen (T200) spans the lipid bilayer and contains a cytoplasmic domain of 80,000 Mr. Cell. 1985 May;41(1):83–93. doi: 10.1016/0092-8674(85)90063-7. [DOI] [PubMed] [Google Scholar]
  34. Trowbridge I. S. Interspecies spleen-myeloma hybrid producing monoclonal antibodies against mouse lymphocyte surface glycoprotein, T200. J Exp Med. 1978 Jul 1;148(1):313–323. doi: 10.1084/jem.148.1.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Trowbridge I. S., Ralph P., Bevan M. J. Differences in the surface proteins of mouse B and T cells. Proc Natl Acad Sci U S A. 1975 Jan;72(1):157–161. doi: 10.1073/pnas.72.1.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Woollett G. R., Barclay A. N., Puklavec M., Williams A. F. Molecular and antigenic heterogeneity of the rat leukocyte-common antigen from thymocytes and T and B lymphocytes. Eur J Immunol. 1985 Feb;15(2):168–173. doi: 10.1002/eji.1830150211. [DOI] [PubMed] [Google Scholar]
  37. Woollett G. R., Williams A. F., Shotton D. M. Visualisation by low-angle shadowing of the leucocyte-common antigen. A major cell surface glycoprotein of lymphocytes. EMBO J. 1985 Nov;4(11):2827–2830. doi: 10.1002/j.1460-2075.1985.tb04010.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zamoyska R., Vollmer A. C., Sizer K. C., Liaw C. W., Parnes J. R. Two Lyt-2 polypeptides arise from a single gene by alternative splicing patterns of mRNA. Cell. 1985 Nov;43(1):153–163. doi: 10.1016/0092-8674(85)90020-0. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES