Aging-dependent proteolysis of NF-κB in human fibroblasts

Tetsuro Ikebe, Eijiro Jimi, Mahiro Beppu, Hiroshi Takeuchi, Hideki Nakayama, Kanemitsu Shirasuna

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

We investigated the NF-κB-like factor induced in the late-passage human oral mucosal fibroblasts stimulated with interleukin-1 (IL-1). Compared with the NFκBs of HeLa cells and early-passage fibroblasts, the NF-κB-like factor of late-passage (passage 15) fibroblasts migrated faster in the electrophoretic mobility shift assay (EMSA) and behaved like a 70-80 kDa protein in the gel filtration chromatography. Both antibodies against p50 and p65 subunits of NF-κB could supershift the small NF-κB-like factor of late- passage cells in the EMSAs. A 47-kDa band was detected in late-passage fibroblasts by immunoblotting against p50. The mobility of the trypsin- degraded NF-κB of HeLa cells corresponded to that of the small NF-κB-like factor of late-passage fibroblasts in the EMSAs. Furthermore, when the nuclear extracts of the IL-1-stimulated HeLa cells were incubated with those of the IL-1-stimulated old fibroblasts, the p65-p50 NF-κB band disappeared, leaving behind a small NF-κB-like band. This reduction of NF-κB was prevented by the addition of a cysteine protease inhibitor leupeptin. These results suggest that the small NF-κB-like factor of late-passage fibroblasts is a part of the NF-κB truncated by aging-induced protease(s).

Original languageEnglish
Pages (from-to)247-255
Number of pages9
JournalJournal of cellular physiology
Volume182
Issue number2
DOIs
Publication statusPublished - Jan 13 2000
Externally publishedYes

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Proteolysis
Fibroblasts
Aging of materials
Interleukin-1
HeLa Cells
Cysteine Proteinase Inhibitors
Electrophoretic mobility
Electrophoretic Mobility Shift Assay
Chromatography
Immunoblotting
Trypsin
Gel Chromatography
Assays
Peptide Hydrolases
Gels
3'-(1-butylphosphoryl)adenosine
Antibodies

All Science Journal Classification (ASJC) codes

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Aging-dependent proteolysis of NF-κB in human fibroblasts. / Ikebe, Tetsuro; Jimi, Eijiro; Beppu, Mahiro; Takeuchi, Hiroshi; Nakayama, Hideki; Shirasuna, Kanemitsu.

In: Journal of cellular physiology, Vol. 182, No. 2, 13.01.2000, p. 247-255.

Research output: Contribution to journalArticle

Ikebe, Tetsuro ; Jimi, Eijiro ; Beppu, Mahiro ; Takeuchi, Hiroshi ; Nakayama, Hideki ; Shirasuna, Kanemitsu. / Aging-dependent proteolysis of NF-κB in human fibroblasts. In: Journal of cellular physiology. 2000 ; Vol. 182, No. 2. pp. 247-255.
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