Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1988 Oct;85(20):7709–7713. doi: 10.1073/pnas.85.20.7709

Identification of specific residues of human interleukin 2 that affect binding to the 70-kDa subunit (p70) of the interleukin 2 receptor.

L Collins 1, W H Tsien 1, C Seals 1, J Hakimi 1, D Weber 1, P Bailon 1, J Hoskings 1, W C Greene 1, V Toome 1, G Ju 1
PMCID: PMC282262  PMID: 3051003

Abstract

Analogs of interleukin 2 containing defined amino acid substitutions and deletions were assayed for bioactivity and for competitive binding to the high-affinity human interleukin 2 receptor complex and its two component subunits, a 55-kDa subunit (p55 or TAC) and a 70-kDa subunit (p70). Substitution of Asp20 or deletion of Phe124 resulted in inactive analog proteins that were unable to interact with the high-affinity p55/p70 complex or the intermediate-affinity p70 subunit of the interleukin 2 receptor. These analogs, however, retained the capacity to compete for binding to the low-affinity p55 subunit. The presence of the carboxylic acid in the side chain of Asp20 was necessary for effective binding to the p70 protein. In contrast, substitution of Trp121 and Leu17 created analogs that were inactive in the bioassay and all three binding assays. The effects of these mutations on protein conformation were assessed by circular dichroism. These results demonstrate that specific residues in the NH2 and COOH termini of interleukin 2 are crucial for its structure and activity.

Full text

PDF
7709

Selected References

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

  1. Arakawa T., Boone T., Davis J. M., Kenney W. C. Structure of unfolded and refolded recombinant derived [Ala125]interleukin 2. Biochemistry. 1986 Dec 16;25(25):8274–8277. doi: 10.1021/bi00373a022. [DOI] [PubMed] [Google Scholar]
  2. Bernard H. U., Helinski D. R. Use of the lambda phage promoter PL to promote gene expression in hybrid plasmid cloning vehicles. Methods Enzymol. 1979;68:482–492. doi: 10.1016/0076-6879(79)68037-0. [DOI] [PubMed] [Google Scholar]
  3. Brandhuber B. J., Boone T., Kenney W. C., McKay D. B. Three-dimensional structure of interleukin-2. Science. 1987 Dec 18;238(4834):1707–1709. doi: 10.1126/science.3500515. [DOI] [PubMed] [Google Scholar]
  4. Dukovich M., Wano Y., Le thi Bich Thuy, Katz P., Cullen B. R., Kehrl J. H., Greene W. C. A second human interleukin-2 binding protein that may be a component of high-affinity interleukin-2 receptors. Nature. 1987 Jun 11;327(6122):518–522. doi: 10.1038/327518a0. [DOI] [PubMed] [Google Scholar]
  5. Greene W. C., Robb R. J., Svetlik P. B., Rusk C. M., Depper J. M., Leonard W. J. Stable expression of cDNA encoding the human interleukin 2 receptor in eukaryotic cells. J Exp Med. 1985 Jul 1;162(1):363–368. doi: 10.1084/jem.162.1.363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hakimi J., Seals C., Anderson L. E., Podlaski F. J., Lin P., Danho W., Jenson J. C., Perkins A., Donadio P. E., Familletti P. C. Biochemical and functional analysis of soluble human interleukin-2 receptor produced in rodent cells. Solid-phase reconstitution of a receptor-ligand binding reaction. J Biol Chem. 1987 Dec 25;262(36):17336–17341. [PubMed] [Google Scholar]
  7. Hatakeyama M., Minamoto S., Uchiyama T., Hardy R. R., Yamada G., Taniguchi T. Reconstitution of functional receptor for human interleukin-2 in mouse cells. Nature. 1985 Dec 5;318(6045):467–470. doi: 10.1038/318467a0. [DOI] [PubMed] [Google Scholar]
  8. Ju G., Collins L., Kaffka K. L., Tsien W. H., Chizzonite R., Crowl R., Bhatt R., Kilian P. L. Structure-function analysis of human interleukin-2. Identification of amino acid residues required for biological activity. J Biol Chem. 1987 Apr 25;262(12):5723–5731. [PubMed] [Google Scholar]
  9. Kuo L. M., Robb R. J. Structure-function relationships for the IL 2-receptor system. I. Localization of a receptor binding site on IL 2. J Immunol. 1986 Sep 1;137(5):1538–1543. [PubMed] [Google Scholar]
  10. Liang S. M., Lee N., Zoon K. C., Manischewitz J. F., Chollet A., Liang C. M., Quinnan G. V. Biological characterization of human interleukin-2 mutant proteins. Structure-activity relationship studies. J Biol Chem. 1988 Apr 5;263(10):4768–4772. [PubMed] [Google Scholar]
  11. Liang S. M., Thatcher D. R., Liang C. M., Allet B. Studies of structure-activity relationships of human interleukin-2. J Biol Chem. 1986 Jan 5;261(1):334–337. [PubMed] [Google Scholar]
  12. Robb R. J., Greene W. C. Internalization of interleukin 2 is mediated by the beta chain of the high-affinity interleukin 2 receptor. J Exp Med. 1987 Apr 1;165(4):1201–1206. doi: 10.1084/jem.165.4.1201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Robb R. J., Greene W. C., Rusk C. M. Low and high affinity cellular receptors for interleukin 2. Implications for the level of Tac antigen. J Exp Med. 1984 Oct 1;160(4):1126–1146. doi: 10.1084/jem.160.4.1126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Robb R. J., Kutny R. M., Panico M., Morris H. R., Chowdhry V. Amino acid sequence and post-translational modification of human interleukin 2. Proc Natl Acad Sci U S A. 1984 Oct;81(20):6486–6490. doi: 10.1073/pnas.81.20.6486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Robb R. J., Munck A., Smith K. A. T cell growth factor receptors. Quantitation, specificity, and biological relevance. J Exp Med. 1981 Nov 1;154(5):1455–1474. doi: 10.1084/jem.154.5.1455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Robb R. J., Rusk C. M., Yodoi J., Greene W. C. Interleukin 2 binding molecule distinct from the Tac protein: analysis of its role in formation of high-affinity receptors. Proc Natl Acad Sci U S A. 1987 Apr;84(7):2002–2006. doi: 10.1073/pnas.84.7.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Siegel J. P., Sharon M., Smith P. L., Leonard W. J. The IL-2 receptor beta chain (p70): role in mediating signals for LAK, NK, and proliferative activities. Science. 1987 Oct 2;238(4823):75–78. doi: 10.1126/science.3116668. [DOI] [PubMed] [Google Scholar]
  18. Tsudo M., Goldman C. K., Bongiovanni K. F., Chan W. C., Winton E. F., Yagita M., Grimm E. A., Waldmann T. A. The p75 peptide is the receptor for interleukin 2 expressed on large granular lymphocytes and is responsible for the interleukin 2 activation of these cells. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5394–5398. doi: 10.1073/pnas.84.15.5394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Wang H. M., Smith K. A. The interleukin 2 receptor. Functional consequences of its bimolecular structure. J Exp Med. 1987 Oct 1;166(4):1055–1069. doi: 10.1084/jem.166.4.1055. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES