Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1995 Dec 1;182(6):1865–1870. doi: 10.1084/jem.182.6.1865

The cleavage preference of the proteasome governs the yield of antigenic peptides

PMCID: PMC2192259  PMID: 7500032

Abstract

Proteasomes degrade endogenous proteins in the cytosol. The potential contribution of the proteasome to the effect of flanking sequences on the presentation of an antigenic epitope presented by the major histocompatibility complex class I allele Ld was studied. Peptides generated in cells from minigenes coding for peptides of 17- and 19- amino acid length were compared with the in vitro 20S proteasome degradation products of the respective synthetic peptides. The quality of generated peptides was independent of ubiquitination. In vivo and in vitro processing products were indistinguishable with respect to peptide size and abundance. Altering the neighboring sequence substantially improved the yield of the final antigenic nonapeptide by 20S proteasome cleavage. These results suggest that, in addition to the presence of major histocompatibility complex class I allelic motifs, the cleavage preference of the proteasome can define the antigenic potential of a protein.

Full Text

The Full Text of this article is available as a PDF (593.7 KB).

Selected References

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

  1. Boes B., Hengel H., Ruppert T., Multhaup G., Koszinowski U. H., Kloetzel P. M. Interferon gamma stimulation modulates the proteolytic activity and cleavage site preference of 20S mouse proteasomes. J Exp Med. 1994 Mar 1;179(3):901–909. doi: 10.1084/jem.179.3.901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chimini G., Pala P., Sire J., Jordan B. R., Maryanski J. L. Recognition of oligonucleotide-encoded T cell epitopes introduced into a gene unrelated to the original antigen. J Exp Med. 1989 Jan 1;169(1):297–302. doi: 10.1084/jem.169.1.297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cox J. H., Galardy P., Bennink J. R., Yewdell J. W. Presentation of endogenous and exogenous antigens is not affected by inactivation of E1 ubiquitin-activating enzyme in temperature-sensitive cell lines. J Immunol. 1995 Jan 15;154(2):511–519. [PubMed] [Google Scholar]
  4. Del Val M., Schlicht H. J., Ruppert T., Reddehase M. J., Koszinowski U. H. Efficient processing of an antigenic sequence for presentation by MHC class I molecules depends on its neighboring residues in the protein. Cell. 1991 Sep 20;66(6):1145–1153. doi: 10.1016/0092-8674(91)90037-y. [DOI] [PubMed] [Google Scholar]
  5. Dick L. R., Aldrich C., Jameson S. C., Moomaw C. R., Pramanik B. C., Doyle C. K., DeMartino G. N., Bevan M. J., Forman J. M., Slaughter C. A. Proteolytic processing of ovalbumin and beta-galactosidase by the proteasome to a yield antigenic peptides. J Immunol. 1994 Apr 15;152(8):3884–3894. [PMC free article] [PubMed] [Google Scholar]
  6. Dick L. R., Moomaw C. R., DeMartino G. N., Slaughter C. A. Degradation of oxidized insulin B chain by the multiproteinase complex macropain (proteasome). Biochemistry. 1991 Mar 12;30(10):2725–2734. doi: 10.1021/bi00224a022. [DOI] [PubMed] [Google Scholar]
  7. Djaballah H., Rivett A. J. Peptidylglutamyl-peptide hydrolase activity of the multicatalytic proteinase complex: evidence for a new high-affinity site, analysis of cooperative kinetics, and the effect of manganese ions. Biochemistry. 1992 Apr 28;31(16):4133–4141. doi: 10.1021/bi00131a033. [DOI] [PubMed] [Google Scholar]
  8. Driscoll J., Brown M. G., Finley D., Monaco J. J. MHC-linked LMP gene products specifically alter peptidase activities of the proteasome. Nature. 1993 Sep 16;365(6443):262–264. doi: 10.1038/365262a0. [DOI] [PubMed] [Google Scholar]
  9. Eisenlohr L. C., Yewdell J. W., Bennink J. R. Flanking sequences influence the presentation of an endogenously synthesized peptide to cytotoxic T lymphocytes. J Exp Med. 1992 Feb 1;175(2):481–487. doi: 10.1084/jem.175.2.481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Eytan E., Ganoth D., Armon T., Hershko A. ATP-dependent incorporation of 20S protease into the 26S complex that degrades proteins conjugated to ubiquitin. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7751–7755. doi: 10.1073/pnas.86.20.7751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fehling H. J., Swat W., Laplace C., Kühn R., Rajewsky K., Müller U., von Boehmer H. MHC class I expression in mice lacking the proteasome subunit LMP-7. Science. 1994 Aug 26;265(5176):1234–1237. doi: 10.1126/science.8066463. [DOI] [PubMed] [Google Scholar]
  12. Frentzel S., Gräf U., Hämmerling G. J., Kloetzel P. M. Isolation and characterization of the MHC linked beta-type proteasome subunit MC13 cDNA. FEBS Lett. 1992 May 11;302(2):121–125. doi: 10.1016/0014-5793(92)80420-l. [DOI] [PubMed] [Google Scholar]
  13. Gaczynska M., Rock K. L., Goldberg A. L. Gamma-interferon and expression of MHC genes regulate peptide hydrolysis by proteasomes. Nature. 1993 Sep 16;365(6443):264–267. doi: 10.1038/365264a0. [DOI] [PubMed] [Google Scholar]
  14. Glynne R., Powis S. H., Beck S., Kelly A., Kerr L. A., Trowsdale J. A proteasome-related gene between the two ABC transporter loci in the class II region of the human MHC. Nature. 1991 Sep 26;353(6342):357–360. doi: 10.1038/353357a0. [DOI] [PubMed] [Google Scholar]
  15. Henderson R. A., Michel H., Sakaguchi K., Shabanowitz J., Appella E., Hunt D. F., Engelhard V. H. HLA-A2.1-associated peptides from a mutant cell line: a second pathway of antigen presentation. Science. 1992 Mar 6;255(5049):1264–1266. doi: 10.1126/science.1546329. [DOI] [PubMed] [Google Scholar]
  16. Hoffman L., Pratt G., Rechsteiner M. Multiple forms of the 20 S multicatalytic and the 26 S ubiquitin/ATP-dependent proteases from rabbit reticulocyte lysate. J Biol Chem. 1992 Nov 5;267(31):22362–22368. [PubMed] [Google Scholar]
  17. Johnson E. S., Bartel B., Seufert W., Varshavsky A. Ubiquitin as a degradation signal. EMBO J. 1992 Feb;11(2):497–505. doi: 10.1002/j.1460-2075.1992.tb05080.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kelly A., Powis S. H., Glynne R., Radley E., Beck S., Trowsdale J. Second proteasome-related gene in the human MHC class II region. Nature. 1991 Oct 17;353(6345):667–668. doi: 10.1038/353667a0. [DOI] [PubMed] [Google Scholar]
  19. Michalek M. T., Grant E. P., Gramm C., Goldberg A. L., Rock K. L. A role for the ubiquitin-dependent proteolytic pathway in MHC class I-restricted antigen presentation. Nature. 1993 Jun 10;363(6429):552–554. doi: 10.1038/363552a0. [DOI] [PubMed] [Google Scholar]
  20. Orlowski M., Cardozo C., Michaud C. Evidence for the presence of five distinct proteolytic components in the pituitary multicatalytic proteinase complex. Properties of two components cleaving bonds on the carboxyl side of branched chain and small neutral amino acids. Biochemistry. 1993 Feb 16;32(6):1563–1572. doi: 10.1021/bi00057a022. [DOI] [PubMed] [Google Scholar]
  21. Ortiz-Navarrete V., Seelig A., Gernold M., Frentzel S., Kloetzel P. M., Hämmerling G. J. Subunit of the '20S' proteasome (multicatalytic proteinase) encoded by the major histocompatibility complex. Nature. 1991 Oct 17;353(6345):662–664. doi: 10.1038/353662a0. [DOI] [PubMed] [Google Scholar]
  22. Reddehase M. J., Rothbard J. B., Koszinowski U. H. A pentapeptide as minimal antigenic determinant for MHC class I-restricted T lymphocytes. Nature. 1989 Feb 16;337(6208):651–653. doi: 10.1038/337651a0. [DOI] [PubMed] [Google Scholar]
  23. Rock K. L., Gramm C., Rothstein L., Clark K., Stein R., Dick L., Hwang D., Goldberg A. L. Inhibitors of the proteasome block the degradation of most cell proteins and the generation of peptides presented on MHC class I molecules. Cell. 1994 Sep 9;78(5):761–771. doi: 10.1016/s0092-8674(94)90462-6. [DOI] [PubMed] [Google Scholar]
  24. Rötzschke O., Falk K., Deres K., Schild H., Norda M., Metzger J., Jung G., Rammensee H. G. Isolation and analysis of naturally processed viral peptides as recognized by cytotoxic T cells. Nature. 1990 Nov 15;348(6298):252–254. doi: 10.1038/348252a0. [DOI] [PubMed] [Google Scholar]
  25. Rötzschke O., Falk K., Wallny H. J., Faath S., Rammensee H. G. Characterization of naturally occurring minor histocompatibility peptides including H-4 and H-Y. Science. 1990 Jul 20;249(4966):283–287. doi: 10.1126/science.1695760. [DOI] [PubMed] [Google Scholar]
  26. Sherman L. A., Burke T. A., Biggs J. A. Extracellular processing of peptide antigens that bind class I major histocompatibility molecules. J Exp Med. 1992 May 1;175(5):1221–1226. doi: 10.1084/jem.175.5.1221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Townsend A., Bastin J., Gould K., Brownlee G., Andrew M., Coupar B., Boyle D., Chan S., Smith G. Defective presentation to class I-restricted cytotoxic T lymphocytes in vaccinia-infected cells is overcome by enhanced degradation of antigen. J Exp Med. 1988 Oct 1;168(4):1211–1224. doi: 10.1084/jem.168.4.1211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wenzel T., Eckerskorn C., Lottspeich F., Baumeister W. Existence of a molecular ruler in proteasomes suggested by analysis of degradation products. FEBS Lett. 1994 Aug 1;349(2):205–209. doi: 10.1016/0014-5793(94)00665-2. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES