Skip to main content
Protein Science : A Publication of the Protein Society logoLink to Protein Science : A Publication of the Protein Society
. 1997 Nov;6(11):2469–2473. doi: 10.1002/pro.5560061123

The proteasome 11S regulator subunit REG alpha (PA28 alpha) is a heptamer.

S C Johnston 1, F G Whitby 1, C Realini 1, M Rechsteiner 1, C P Hill 1
PMCID: PMC2143584  PMID: 9385652

Abstract

Activity of the 20S proteasome, which performs much of the cytosolic and nuclear proteolysis in eukaryotic cells, is controlled by regulatory complexes that bind to one or both ends of the cylindrical proteasome. One of these complexes, the 11S regulator (REG), is a complex of 28 kDa subunits that is thought to activate proteasomes toward the production of antigenic peptides. REG, purified from red blood cells, is a complex of REG alpha and REG beta subunits. We have crystallized recombinant REG alpha (rREG alpha) and collected diffraction data to 3.0 A resolution. The self-rotation function indicates that rREG alpha forms a heptameric ring in the crystal. Equilibrium sedimentation demonstrates that rREG alpha is a heptamer in solution also.

Full Text

The Full Text of this article is available as a PDF (487.7 KB).

Selected References

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

  1. Ahn J. Y., Tanahashi N., Akiyama K., Hisamatsu H., Noda C., Tanaka K., Chung C. H., Shibmara N., Willy P. J., Mott J. D. Primary structures of two homologous subunits of PA28, a gamma-interferon-inducible protein activator of the 20S proteasome. FEBS Lett. 1995 Jun 5;366(1):37–42. doi: 10.1016/0014-5793(95)00492-r. [DOI] [PubMed] [Google Scholar]
  2. Ahn K., Erlander M., Leturcq D., Peterson P. A., Früh K., Yang Y. In vivo characterization of the proteasome regulator PA28. J Biol Chem. 1996 Jul 26;271(30):18237–18242. doi: 10.1074/jbc.271.30.18237. [DOI] [PubMed] [Google Scholar]
  3. Brooks I., Watts D. G., Soneson K. K., Hensley P. Determining confidence intervals for parameters derived from analysis of equilibrium analytical ultracentrifugation data. Methods Enzymol. 1994;240:459–478. doi: 10.1016/s0076-6879(94)40060-1. [DOI] [PubMed] [Google Scholar]
  4. Cardozo C., Vinitsky A., Michaud C., Orlowski M. Evidence that the nature of amino acid residues in the P3 position directs substrates to distinct catalytic sites of the pituitary multicatalytic proteinase complex (proteasome). Biochemistry. 1994 May 31;33(21):6483–6489. doi: 10.1021/bi00187a014. [DOI] [PubMed] [Google Scholar]
  5. Chu-Ping M., Vu J. H., Proske R. J., Slaughter C. A., DeMartino G. N. Identification, purification, and characterization of a high molecular weight, ATP-dependent activator (PA700) of the 20 S proteasome. J Biol Chem. 1994 Feb 4;269(5):3539–3547. [PubMed] [Google Scholar]
  6. DeMartino G. N., Moomaw C. R., Zagnitko O. P., Proske R. J., Chu-Ping M., Afendis S. J., Swaffield J. C., Slaughter C. A. PA700, an ATP-dependent activator of the 20 S proteasome, is an ATPase containing multiple members of a nucleotide-binding protein family. J Biol Chem. 1994 Aug 19;269(33):20878–20884. [PubMed] [Google Scholar]
  7. Di Cola D. Human erythrocyte contains a factor that stimulates the peptidase activities of multicatalytic proteinase complex. Ital J Biochem. 1992 Jul-Aug;41(4):213–224. [PubMed] [Google Scholar]
  8. Dick T. P., Ruppert T., Groettrup M., Kloetzel P. M., Kuehn L., Koszinowski U. H., Stevanović S., Schild H., Rammensee H. G. Coordinated dual cleavages induced by the proteasome regulator PA28 lead to dominant MHC ligands. Cell. 1996 Jul 26;86(2):253–262. doi: 10.1016/s0092-8674(00)80097-5. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Dubiel W., Pratt G., Ferrell K., Rechsteiner M. Purification of an 11 S regulator of the multicatalytic protease. J Biol Chem. 1992 Nov 5;267(31):22369–22377. [PubMed] [Google Scholar]
  11. 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]
  12. Germain R. N. MHC-dependent antigen processing and peptide presentation: providing ligands for T lymphocyte activation. Cell. 1994 Jan 28;76(2):287–299. doi: 10.1016/0092-8674(94)90336-0. [DOI] [PubMed] [Google Scholar]
  13. Gray C. W., Slaughter C. A., DeMartino G. N. PA28 activator protein forms regulatory caps on proteasome stacked rings. J Mol Biol. 1994 Feb 11;236(1):7–15. doi: 10.1006/jmbi.1994.1113. [DOI] [PubMed] [Google Scholar]
  14. Hoffman L., Rechsteiner M. Regulatory features of multicatalytic and 26S proteases. Curr Top Cell Regul. 1996;34:1–32. doi: 10.1016/s0070-2137(96)80001-x. [DOI] [PubMed] [Google Scholar]
  15. Löwe J., Stock D., Jap B., Zwickl P., Baumeister W., Huber R. Crystal structure of the 20S proteasome from the archaeon T. acidophilum at 3.4 A resolution. Science. 1995 Apr 28;268(5210):533–539. doi: 10.1126/science.7725097. [DOI] [PubMed] [Google Scholar]
  16. Ma C. P., Slaughter C. A., DeMartino G. N. Identification, purification, and characterization of a protein activator (PA28) of the 20 S proteasome (macropain). J Biol Chem. 1992 May 25;267(15):10515–10523. [PubMed] [Google Scholar]
  17. Matthews B. W. Solvent content of protein crystals. J Mol Biol. 1968 Apr 28;33(2):491–497. doi: 10.1016/0022-2836(68)90205-2. [DOI] [PubMed] [Google Scholar]
  18. Peters J. M., Cejka Z., Harris J. R., Kleinschmidt J. A., Baumeister W. Structural features of the 26 S proteasome complex. J Mol Biol. 1993 Dec 20;234(4):932–937. doi: 10.1006/jmbi.1993.1646. [DOI] [PubMed] [Google Scholar]
  19. Realini C., Jensen C. C., Zhang Z., Johnston S. C., Knowlton J. R., Hill C. P., Rechsteiner M. Characterization of recombinant REGalpha, REGbeta, and REGgamma proteasome activators. J Biol Chem. 1997 Oct 10;272(41):25483–25492. doi: 10.1074/jbc.272.41.25483. [DOI] [PubMed] [Google Scholar]
  20. Rechsteiner M., Hoffman L., Dubiel W. The multicatalytic and 26 S proteases. J Biol Chem. 1993 Mar 25;268(9):6065–6068. [PubMed] [Google Scholar]
  21. Song X., Mott J. D., von Kampen J., Pramanik B., Tanaka K., Slaughter C. A., DeMartino G. N. A model for the quaternary structure of the proteasome activator PA28. J Biol Chem. 1996 Oct 18;271(42):26410–26417. doi: 10.1074/jbc.271.42.26410. [DOI] [PubMed] [Google Scholar]
  22. Tong L. A., Rossmann M. G. The locked rotation function. Acta Crystallogr A. 1990 Oct 1;46(Pt 10):783–792. doi: 10.1107/s0108767390005530. [DOI] [PubMed] [Google Scholar]
  23. Townsend A., Bodmer H. Antigen recognition by class I-restricted T lymphocytes. Annu Rev Immunol. 1989;7:601–624. doi: 10.1146/annurev.iy.07.040189.003125. [DOI] [PubMed] [Google Scholar]
  24. Ustrell V., Pratt G., Rechsteiner M. Effects of interferon gamma and major histocompatibility complex-encoded subunits on peptidase activities of human multicatalytic proteases. Proc Natl Acad Sci U S A. 1995 Jan 17;92(2):584–588. doi: 10.1073/pnas.92.2.584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wenzel T., Baumeister W. Conformational constraints in protein degradation by the 20S proteasome. Nat Struct Biol. 1995 Mar;2(3):199–204. doi: 10.1038/nsb0395-199. [DOI] [PubMed] [Google Scholar]
  26. Yewdell J. W., Bennink J. R. Cell biology of antigen processing and presentation to major histocompatibility complex class I molecule-restricted T lymphocytes. Adv Immunol. 1992;52:1–123. doi: 10.1016/s0065-2776(08)60875-5. [DOI] [PubMed] [Google Scholar]
  27. Yukawa M., Sakon M., Kambayashi J., Shiba E., Kawasaki T., Ariyoshi H., Mori T. Proteasome and its novel endogeneous activator in human platelets. Biochem Biophys Res Commun. 1991 Jul 15;178(1):256–262. doi: 10.1016/0006-291x(91)91807-o. [DOI] [PubMed] [Google Scholar]

Articles from Protein Science : A Publication of the Protein Society are provided here courtesy of The Protein Society

RESOURCES