Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1990 May 15;268(1):135–140. doi: 10.1042/bj2680135

Nucleotide sequences and three-dimensional modelling of the VH and VL domains of two human monoclonal antibodies specific for the D antigen of the human Rh-blood-group system.

N C Hughes-Jones 1, J M Bye 1, D Beale 1, J Coadwell 1
PMCID: PMC1131402  PMID: 2111699

Abstract

The nucleotide sequences were determined for the VH and VL domains of two human IgG1 antibodies, Pag-1 and Fog-B, specific for the D antigen of the Rh-blood-group system. The VH-region genes of the two antibodies were derived from separate germ-line genes within the VH-IV gene family, but both antibodies used the same JH6 gene. The D-region genes differed from each other, and no similarity was found to known D regions. The light chain of Fog-B belongs to the V lambda-I subgroup and that of Pag-1 probably belongs to the V lambda-V subgroup; both light chains used the J2/3 gene. Three-dimensional models of the variable domains were made, based on those of known crystallographic structure. The surface contours at the combining sites are clearly different, consistent with the evidence that the antibodies recognize different but overlapping epitopes. Some details of the molecular modelling of hypervariable regions have been deposited as Supplementary Publication SUP 50155 (6 pages) at the British Library Document Supply Centre, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1990) 265, 5.

Full text

PDF
135

Selected References

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

  1. Amit A. G., Mariuzza R. A., Phillips S. E., Poljak R. J. Three-dimensional structure of an antigen-antibody complex at 2.8 A resolution. Science. 1986 Aug 15;233(4765):747–753. doi: 10.1126/science.2426778. [DOI] [PubMed] [Google Scholar]
  2. Berek C., Griffiths G. M., Milstein C. Molecular events during maturation of the immune response to oxazolone. Nature. 1985 Aug 1;316(6027):412–418. doi: 10.1038/316412a0. [DOI] [PubMed] [Google Scholar]
  3. Bernstein F. C., Koetzle T. F., Williams G. J., Meyer E. F., Jr, Brice M. D., Rodgers J. R., Kennard O., Shimanouchi T., Tasumi M. The Protein Data Bank: a computer-based archival file for macromolecular structures. J Mol Biol. 1977 May 25;112(3):535–542. doi: 10.1016/s0022-2836(77)80200-3. [DOI] [PubMed] [Google Scholar]
  4. Buluwela L., Albertson D. G., Sherrington P., Rabbitts P. H., Spurr N., Rabbitts T. H. The use of chromosomal translocations to study human immunoglobulin gene organization: mapping DH segments within 35 kb of the C mu gene and identification of a new DH locus. EMBO J. 1988 Jul;7(7):2003–2010. doi: 10.1002/j.1460-2075.1988.tb03039.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chothia C., Lesk A. M. Canonical structures for the hypervariable regions of immunoglobulins. J Mol Biol. 1987 Aug 20;196(4):901–917. doi: 10.1016/0022-2836(87)90412-8. [DOI] [PubMed] [Google Scholar]
  6. Chothia C., Lesk A. M., Levitt M., Amit A. G., Mariuzza R. A., Phillips S. E., Poljak R. J. The predicted structure of immunoglobulin D1.3 and its comparison with the crystal structure. Science. 1986 Aug 15;233(4765):755–758. doi: 10.1126/science.3090684. [DOI] [PubMed] [Google Scholar]
  7. Clarke S. H., Huppi K., Ruezinsky D., Staudt L., Gerhard W., Weigert M. Inter- and intraclonal diversity in the antibody response to influenza hemagglutinin. J Exp Med. 1985 Apr 1;161(4):687–704. doi: 10.1084/jem.161.4.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Crews S., Griffin J., Huang H., Calame K., Hood L. A single VH gene segment encodes the immune response to phosphorylcholine: somatic mutation is correlated with the class of the antibody. Cell. 1981 Jul;25(1):59–66. doi: 10.1016/0092-8674(81)90231-2. [DOI] [PubMed] [Google Scholar]
  9. Dariavach P., Lefranc G., Lefranc M. P. Human immunoglobulin C lambda 6 gene encodes the Kern+Oz-lambda chain and C lambda 4 and C lambda 5 are pseudogenes. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9074–9078. doi: 10.1073/pnas.84.24.9074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Epp O., Lattman E. E., Schiffer M., Huber R., Palm W. The molecular structure of a dimer composed of the variable portions of the Bence-Jones protein REI refined at 2.0-A resolution. Biochemistry. 1975 Nov 4;14(22):4943–4952. doi: 10.1021/bi00693a025. [DOI] [PubMed] [Google Scholar]
  11. Eulitz M. A new subgroup of human L-chains of the lambda-type. Primary structure of Bence-Jones protein DEL. Eur J Biochem. 1974 Dec 16;50(1):49–69. doi: 10.1111/j.1432-1033.1974.tb03872.x. [DOI] [PubMed] [Google Scholar]
  12. Favaloro J., Treisman R., Kamen R. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980;65(1):718–749. doi: 10.1016/s0076-6879(80)65070-8. [DOI] [PubMed] [Google Scholar]
  13. Furey W., Jr, Wang B. C., Yoo C. S., Sax M. Structure of a novel Bence-Jones protein (Rhe) fragment at 1.6 A resolution. J Mol Biol. 1983 Jul 5;167(3):661–692. doi: 10.1016/s0022-2836(83)80104-1. [DOI] [PubMed] [Google Scholar]
  14. Gorick B. D., Thompson K. M., Melamed M. D., Hughes-Jones N. C. Three epitopes on the human Rh antigen D recognized by 125I-labelled human monoclonal IgG antibodies. Vox Sang. 1988;55(3):165–170. doi: 10.1111/j.1423-0410.1988.tb05086.x. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. HUGHES-JONES N. C., GARDNER B., TELFORD R. THE EFFECT OF PH AND IONIC STRENGTH ON THE REACTION BETWEEN ANTI-D AND ERYTHROCYTES. Immunology. 1964 Jan;7:72–81. [PMC free article] [PubMed] [Google Scholar]
  17. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  18. Ichihara Y., Abe M., Yasui H., Matsuoka H., Kurosawa Y. At least five DH genes of human immunoglobulin heavy chains are encoded in 9-kilobase DNA fragments. Eur J Immunol. 1988 Apr;18(4):649–652. doi: 10.1002/eji.1830180426. [DOI] [PubMed] [Google Scholar]
  19. Kishimoto T., Okajima H., Okumoto T., Taniguchi M. Nucleotide sequences of the cDNAs encoding the V-regions of H- and L-chains of a human monoclonal antibody with broad reactivity to malignant tumor cells. Nucleic Acids Res. 1989 Jun 12;17(11):4385–4385. doi: 10.1093/nar/17.11.4385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lascombe M. B., Alzari P. M., Boulot G., Saludjian P., Tougard P., Berek C., Haba S., Rosen E. M., Nisonoff A., Poljak R. J. Three-dimensional structure of Fab R19.9, a monoclonal murine antibody specific for the p-azobenzenearsonate group. Proc Natl Acad Sci U S A. 1989 Jan;86(2):607–611. doi: 10.1073/pnas.86.2.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lee K. H., Matsuda F., Kinashi T., Kodaira M., Honjo T. A novel family of variable region genes of the human immunoglobulin heavy chain. J Mol Biol. 1987 Jun 20;195(4):761–768. doi: 10.1016/0022-2836(87)90482-7. [DOI] [PubMed] [Google Scholar]
  22. Lomas C., Tippett P., Thompson K. M., Melamed M. D., Hughes-Jones N. C. Demonstration of seven epitopes on the Rh antigen D using human monoclonal anti-D antibodies and red cells from D categories. Vox Sang. 1989;57(4):261–264. doi: 10.1111/j.1423-0410.1989.tb00839.x. [DOI] [PubMed] [Google Scholar]
  23. Marquart M., Deisenhofer J., Huber R., Palm W. Crystallographic refinement and atomic models of the intact immunoglobulin molecule Kol and its antigen-binding fragment at 3.0 A and 1.0 A resolution. J Mol Biol. 1980 Aug 25;141(4):369–391. doi: 10.1016/0022-2836(80)90252-1. [DOI] [PubMed] [Google Scholar]
  24. McGregor M. J., Islam S. A., Sternberg M. J. Analysis of the relationship between side-chain conformation and secondary structure in globular proteins. J Mol Biol. 1987 Nov 20;198(2):295–310. doi: 10.1016/0022-2836(87)90314-7. [DOI] [PubMed] [Google Scholar]
  25. Melamed M. D., Thompson K. M., Gibson T., Hughes-Jones N. C. Requirements for the establishment of heterohybridomas secreting monoclonal human antibody to rhesus (D) blood group antigen. J Immunol Methods. 1987 Nov 23;104(1-2):245–251. doi: 10.1016/0022-1759(87)90511-4. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Mudgett-Hunter M., Anderson W., Haber E., Margolies M. N. Binding and structural diversity among high-affinity monoclonal anti-digoxin antibodies. Mol Immunol. 1985 Apr;22(4):477–488. doi: 10.1016/0161-5890(85)90132-4. [DOI] [PubMed] [Google Scholar]
  29. Padlan E. A., Kabat E. A. Model-building study of the combining sites of two antibodies to alpha (1----6)dextran. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6885–6889. doi: 10.1073/pnas.85.18.6885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Padlan E. A., Silverton E. W., Sheriff S., Cohen G. H., Smith-Gill S. J., Davies D. R. Structure of an antibody-antigen complex: crystal structure of the HyHEL-10 Fab-lysozyme complex. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5938–5942. doi: 10.1073/pnas.86.15.5938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ponder J. W., Richards F. M. Tertiary templates for proteins. Use of packing criteria in the enumeration of allowed sequences for different structural classes. J Mol Biol. 1987 Feb 20;193(4):775–791. doi: 10.1016/0022-2836(87)90358-5. [DOI] [PubMed] [Google Scholar]
  32. Ravetch J. V., Siebenlist U., Korsmeyer S., Waldmann T., Leder P. Structure of the human immunoglobulin mu locus: characterization of embryonic and rearranged J and D genes. Cell. 1981 Dec;27(3 Pt 2):583–591. doi: 10.1016/0092-8674(81)90400-1. [DOI] [PubMed] [Google Scholar]
  33. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sanz I., Capra J. D. V kappa and J kappa gene segments of A/J Ars-A antibodies: somatic recombination generates the essential arginine at the junction of the variable and joining regions. Proc Natl Acad Sci U S A. 1987 Feb;84(4):1085–1089. doi: 10.1073/pnas.84.4.1085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Saul F. A., Amzel L. M., Poljak R. J. Preliminary refinement and structural analysis of the Fab fragment from human immunoglobulin new at 2.0 A resolution. J Biol Chem. 1978 Jan 25;253(2):585–597. [PubMed] [Google Scholar]
  36. Segal D. M., Padlan E. A., Cohen G. H., Rudikoff S., Potter M., Davies D. R. The three-dimensional structure of a phosphorylcholine-binding mouse immunoglobulin Fab and the nature of the antigen binding site. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4298–4302. doi: 10.1073/pnas.71.11.4298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sheriff S., Silverton E. W., Padlan E. A., Cohen G. H., Smith-Gill S. J., Finzel B. C., Davies D. R. Three-dimensional structure of an antibody-antigen complex. Proc Natl Acad Sci U S A. 1987 Nov;84(22):8075–8079. doi: 10.1073/pnas.84.22.8075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sibanda B. L., Blundell T. L., Thornton J. M. Conformation of beta-hairpins in protein structures. A systematic classification with applications to modelling by homology, electron density fitting and protein engineering. J Mol Biol. 1989 Apr 20;206(4):759–777. doi: 10.1016/0022-2836(89)90583-4. [DOI] [PubMed] [Google Scholar]
  39. Siebenlist U., Ravetch J. V., Korsmeyer S., Waldmann T., Leder P. Human immunoglobulin D segments encoded in tandem multigenic families. Nature. 1981 Dec 17;294(5842):631–635. doi: 10.1038/294631a0. [DOI] [PubMed] [Google Scholar]
  40. Suh S. W., Bhat T. N., Navia M. A., Cohen G. H., Rao D. N., Rudikoff S., Davies D. R. The galactan-binding immunoglobulin Fab J539: an X-ray diffraction study at 2.6-A resolution. Proteins. 1986 Sep;1(1):74–80. doi: 10.1002/prot.340010112. [DOI] [PubMed] [Google Scholar]
  41. Thompson K. M., Hough D. W., Maddison P. J., Melamed M. D., Hughes-Jones N. The efficient production of stable, human monoclonal antibody-secreting hybridomas from EBV-transformed lymphocytes using the mouse myeloma X63-Ag8.653 as a fusion partner. J Immunol Methods. 1986 Nov 20;94(1-2):7–12. doi: 10.1016/0022-1759(86)90208-5. [DOI] [PubMed] [Google Scholar]
  42. Udey J. A., Blomberg B. Human lambda light chain locus: organization and DNA sequences of three genomic J regions. Immunogenetics. 1987;25(1):63–70. doi: 10.1007/BF00768834. [DOI] [PubMed] [Google Scholar]
  43. Wilmot C. M., Thornton J. M. Analysis and prediction of the different types of beta-turn in proteins. J Mol Biol. 1988 Sep 5;203(1):221–232. doi: 10.1016/0022-2836(88)90103-9. [DOI] [PubMed] [Google Scholar]
  44. Wilson P. W., Rogers J., Harding M., Pohl V., Pattyn G., Lawson D. E. Structure of chick chromosomal genes for calbindin and calretinin. J Mol Biol. 1988 Apr 20;200(4):615–625. doi: 10.1016/0022-2836(88)90475-5. [DOI] [PubMed] [Google Scholar]
  45. Yamasaki N., Komori S., Watanabe T. Complementary DNA for a human subgroup IV immunoglobulin lambda-chain. Mol Immunol. 1987 Sep;24(9):981–985. doi: 10.1016/0161-5890(87)90010-1. [DOI] [PubMed] [Google Scholar]

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

RESOURCES