The Interferon-Gamma-inducible 11 S Regulator (PA28) and the LMP2/LMP7 Subunits Govern the Peptide Production by the 20 S Proteasome in Vitro

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1995
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Ruppert, Thomas
Kuehn, Lothar
Seeger, Michael
Standera, Sybille
Koszinowski, Ulrich
Kloetzel, Peter-M.
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Journal of Biological Chemistry. 1995, 270(40), pp. 23808-23815. ISSN 0021-9258. eISSN 1083-351X. Available under: doi: 10.1074/jbc.270.40.23808
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Antigenic peptides presented on major histocompatibility complex (MHC) class I molecules to cytotoxic T cells are generated in the cytosol by the 20 S proteasome. Upon stimulation of antigen presenting cells with interferon-gamma, two constitutive subunits of the 20 S proteasome are replaced by the MHC-encoded subunits low molecular mass polypeptide (LMP) 2 and LMP 7. In addition the expression of the two subunits of the 11 S regulator of the 20 S proteasome (PA28) are increased. As the function of LMP2 and LMP7 in antigen presentation is still controversial, we tested whether these subunits might operate by modifying proteasome activation through the 11 S regulator. We strongly overexpressed the two LMP subunits separately or together by transfection in murine fibroblasts. Isolated 20 S proteasomes from LMP transfectants were applied in digests of a 25-mer peptide in the presence or absence of a purified preparation of 11 S regulator from rabbit erythrocytes. Analysis of the cleavage products by high performance liquid chromatography and electrospray mass spectroscopy revealed marked differences in the peptide product profile in dependence on the LMP2 and LMP7 content. While the 11 S regulator did not preferentially activate LMP2 or 7 containing proteasomes, the binding of the 11 S regulator to any of the proteasome preparations markedly changed both the quality and quantity of peptides produced. These results suggest that the 11 S regulator increases the spectrum of peptides which can be generated in antigen presenting cells.

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570 Biowissenschaften, Biologie
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ISO 690GRÖTTRUP, Marcus, Thomas RUPPERT, Lothar KUEHN, Michael SEEGER, Sybille STANDERA, Ulrich KOSZINOWSKI, Peter-M. KLOETZEL, 1995. The Interferon-Gamma-inducible 11 S Regulator (PA28) and the LMP2/LMP7 Subunits Govern the Peptide Production by the 20 S Proteasome in Vitro. In: Journal of Biological Chemistry. 1995, 270(40), pp. 23808-23815. ISSN 0021-9258. eISSN 1083-351X. Available under: doi: 10.1074/jbc.270.40.23808
BibTex
@article{Grottrup1995Inter-22321,
  year={1995},
  doi={10.1074/jbc.270.40.23808},
  title={The Interferon-Gamma-inducible 11 S Regulator (PA28) and the LMP2/LMP7 Subunits Govern the Peptide Production by the 20 S Proteasome in Vitro},
  number={40},
  volume={270},
  issn={0021-9258},
  journal={Journal of Biological Chemistry},
  pages={23808--23815},
  author={Gröttrup, Marcus and Ruppert, Thomas and Kuehn, Lothar and Seeger, Michael and Standera, Sybille and Koszinowski, Ulrich and Kloetzel, Peter-M.}
}
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