Abstract
Proteasomes are large multicatalytic proteinases located in the nuclei and cytoplasm of all eukaryotic cells. Proteasomes are composed of four heptameric rings stacked to form a hollow cylinder (length 16-20 nm, diameter 11-12 nm). The outer two rings are composed of α-subunits, while β-subunits, which contain the active sites, comprise the inner two rings. Proteasomes from archaebacteria contain only one type each of α- and β-subunits. Eukaryotic proteasomes are more divergent; yeast proteasomes have seven different α- and seven different β-subunits, each occupying a unique position in the ring. Only three of the seven yeast β-subunits contain the N-terminal threonine necessary for activity (1-2).
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References
Monaco, J. J. and Nandi, D. (1995) Genetics of proteasomes and antigen processing. Annu. Rev. Genetics. 29, 729–754.
Coux, O., Tanaka, K., and Goldberg, A. L. (1996) Structure and functions of the 20S and 26S proteasomes. Annu. Rev. Biochem. 65, 801–847.
Nandi, D., Marusina, K., and Monaco, J. J. (1998) How do endogenous proteins become peptides and reach the endoplasmic reticulum? Curr. Top. Microbiol. Immunol. 232, 15–47.
Griffin, T.A., Nandi, D., Cruz, M., Fehling, H. J., Van Kaer, L., Monaco, J. J., and Colbert, R.A. (1998) Immunoproteasome assembly: cooperative incorporation of interferon γ (IFN-γ)-inducible subunits. J. Exp. Med. 187, 97–104.
Eleuteri, A. M., Kohanski, R. A., Cardozo, C., and Orlowski, M. (1997) Bovine spleen multicatalytic proteinase complex (proteasome): replacement of X, Y, and Z subunits by LMP7, LPM2, and MECL1 and changes in properties and specificity. J. Biol. Chem. 272, 11,824–11,831.
Tanaka, K., Ii, K., Ichihara, A., Waxman, L., and Goldberg, A. L. (1986) A high molecular weight protease in the cytosol of rat liver: purification, enzymological properties, and tissue distribution. J. Biol. Chem. 261, 15,197–15,203.
Salter, R. D., Howell, D. N., and Cresswell, P. (1985) Genes regulating HLA class I antigen expression in T-B lymphoblastoid hybrids. Immunogenetics 21, 235–246.
Brown, M. G. and Monaco, J. J. (1993) Biochemical purification of distinct proteasome subsets. Enzyme Protein 47, 343–353.
Grainger, J. L. and Winkler, M. M. (1989) The sea urchin multicatalytic protease: purification, biochemical analysis, subcellular distribution, and relationship to snRNPs. J. CellBiol. 109, 675–683.
Hua, S.-b., To, W.-Y., Nguyen, T. T., Wong, M.-L., and Wang, C. C. (1996) Purification and characterization of proteasomes from Trypanosoma brucei. Mol. Biochem. Parasitol. 78, 33–46.
Inaba, K., Akazome, Y., and Morisawa, M. (1993) Purification of proteasomes from salmonid fish sperm and their localization along sperm flagella. J. Cell Sci. 104, 907–915.
Klinkradt, S., Naude, R. J., Muramoto, K., and Oelofsen, W. (1997) Purification and characterization of proteasome from ostrich liver. Int. J. Biochem. Cell Biol. 29, 611–622.
Koohmaraie, M. (1992) Ovine skeletal muscle multicatalytic proteinase complex (proteasome): purification, characterization, and comparison of its effects on myofibrils with µ-calpains. J. Anim. Sci. 70, 3697–3708.
Mykles, D. L. (1989) Purification and characterization of a multicatalytic proteinase from crustacean muscle: comparison of latent and heat-activated forms. Arch. Biochem. Biophys. 274, 216–228.
Ozaki, M., Fujinami, K., Tanaka, K., Amemiya, Y., Sato, T., Ogura, N., and Nakagawa, H. (1992) Purification and initial characterization of the proteasome from the higher plant Spinacia oleracea. J. Biol. Chem. 27, 21,678–21,684.
Rivett, A J., Savory, P.J., and Djaballah, H. (1994) Multicatalytic endopeptidase complex: proteasome. Meth. Enzymol. 244, 331–350.
Sacchetta, P., Battista, P., Santarone, S., and Di Cola, D. (1990) Purification of human erythrocyte proteolytic enzyme responsible for degradation of oxidant-damaged hemoglobin. Evidence for identifying as a member of the multicatalytic proteinase family. Biochim. Biophys. Acta. 1037, 337–343.
Saitoh, Y., Yokosawa, H., Takahashi, K., and Ishii, S.-i. (1989) Purification and characterization of multicatalytic proteinase from eggs of the ascidian Halocynthia roretzi. J. Biochem. 105, 254–260.
Suga, Y., Takamori, K., and Ogawa, H. (1993) Skin proteasomes (high-molecular-weight protease): purification, enzymologic properties, gross structure, and tissue distribution. J. Invest. Dermatol. 101, 346–351.
Ma, C.-P., Slaughter, C. A., and DeMartino, G. N. (1992) Identification, purification, and characterization of a protein activator (PA28) of the 20S proteasome (macropain). J. Biol. Chem. 267, 10,515–10,523.
Rivett, A. J. (1993) Proteasomes: multicatalytic proteinase complexes. Biochem. J. 291, 1–10.
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Beyette, J.R., Hubbell, T., Monaco, J.J. (2000). Purification of 20S Proteasomes. In: Solheim, J.C. (eds) Antigen Processing and Presentation Protocols. Methods in Molecular Biology, vol 156. Humana Press. https://doi.org/10.1385/1-59259-062-4:1
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DOI: https://doi.org/10.1385/1-59259-062-4:1
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