Evidence for proteasome dysfunction in cytotoxicity mediated by anti-Ras intracellular antibodies

Alessio Cardinale, Ilaria Filesi, Sonia Mattei, Silvia Biocca

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Anti-Ras intracellular antibodies inhibit cell proliferation in vivo by sequestering the antigen and diverting it from its physiological location [Lener, M., Horn, I. R., Cardinale, A., Messina, S., Nielsen, U.B., Rybak, S.M., Hoogenboom, H.R., Cattaneo, A., Biocca, S. (2000) Eur. J. Biochem. 267, 1196-1205]. Here we demonstrate that strongly aggregating single-chain antibody fragments (scFv), binding to Ras, induce apoptosis, and this effect is strictly related to the antibody-mediated aggregation of p21Ras. Proteasomes are quickly recruited to the newly formed aggregates, and their activity is strongly inhibited. This leads to the formation of aggresome-like structures, which become evident in the vast majority of apoptotic cells. A combination of anti-Ras scFv fragments with a nontoxic concentration of the proteasome inhibitor, lactacystin, markedly increases proteasome dysfunction and apoptosis. The dominant-negative H-ras (N17-H-ras), which is mostly soluble and does not induce aggresome formation or inhibit proteasome activity, only affects cell viability slightly. Together, these observations suggest a mechanism linking antibody-mediated Ras aggregation, impairment of the ubiquitin-proteasome system, and cytotoxicity.

Original languageEnglish
Pages (from-to)3389-3397
Number of pages9
JournalEuropean Journal of Biochemistry
Volume270
Issue number16
DOIs
Publication statusPublished - Aug 2003

Fingerprint

Proteasome Endopeptidase Complex
Cytotoxicity
Antibodies
Agglomeration
Cells
Apoptosis
Single-Chain Antibodies
Immunoglobulin Fragments
Proteasome Inhibitors
Cell proliferation
Horns
Ubiquitin
Cell Survival
Cell Proliferation
Antigens

Keywords

  • Aggresome
  • Anti-p21Ras
  • Apoptosis
  • Proteasome
  • scFv fragment

ASJC Scopus subject areas

  • Biochemistry

Cite this

Evidence for proteasome dysfunction in cytotoxicity mediated by anti-Ras intracellular antibodies. / Cardinale, Alessio; Filesi, Ilaria; Mattei, Sonia; Biocca, Silvia.

In: European Journal of Biochemistry, Vol. 270, No. 16, 08.2003, p. 3389-3397.

Research output: Contribution to journalArticle

Cardinale, Alessio ; Filesi, Ilaria ; Mattei, Sonia ; Biocca, Silvia. / Evidence for proteasome dysfunction in cytotoxicity mediated by anti-Ras intracellular antibodies. In: European Journal of Biochemistry. 2003 ; Vol. 270, No. 16. pp. 3389-3397.
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