Overexpression of the two-chain form of cathepsin B in senescent WI-38 cells

Byron R. Dipaolo, Robert Pignolo, Vincent J. Cristofalo

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

We have examined differential protein expression in serum-stimulated young and senescent WI-38 human fetal lung-derived cells in culture using high-resolution two-dimensional gel electrophoresis. Overexpression of a protein with an approximate Mr of 29,000 and pI of 5.8 was observed in senescent cells during the G0 and throughout the G1 stage of the cell cycle. Automated amino-terminal sequencing of the peptide from polyvinylidene difluoride electroblots showed 100% sequence identity to cathepsin B or pre-procathepsin B in a 12-amino acid overlap, beginning at residue 48 or 129, respectively. The 29-kDa peptide corresponds to the heavy chain of the two-chain enzyme form. Cathepsin B activity was found to be decreased in cells aged in vitro in comparison to that in young controls. Changes in the steady-state levels of both the 4.0- and the 2.2-kb cathespin B transcripts between young and senescent cells cannot account for the overexpression of the two-chain form of the enzyme. These results suggest that increased proteolysis of a conformationally more labile single-chain form and/or decreased turnover and accumulation of a less active form of this lysomosomal protease occur in senescent fibroblasts and may account for the observed decreased cathepsin B activity in senescent cells in culture.

Original languageEnglish (US)
Pages (from-to)500-505
Number of pages6
JournalExperimental Cell Research
Volume201
Issue number2
DOIs
StatePublished - Jan 1 1992
Externally publishedYes

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Cathepsin B
Cell Culture Techniques
Peptides
Electrophoresis, Gel, Two-Dimensional
Enzymes
Proteolysis
Cell Cycle
Proteins
Peptide Hydrolases
Fibroblasts
Amino Acids
Lung
Serum

ASJC Scopus subject areas

  • Cell Biology

Cite this

Overexpression of the two-chain form of cathepsin B in senescent WI-38 cells. / Dipaolo, Byron R.; Pignolo, Robert; Cristofalo, Vincent J.

In: Experimental Cell Research, Vol. 201, No. 2, 01.01.1992, p. 500-505.

Research output: Contribution to journalArticle

Dipaolo, Byron R. ; Pignolo, Robert ; Cristofalo, Vincent J. / Overexpression of the two-chain form of cathepsin B in senescent WI-38 cells. In: Experimental Cell Research. 1992 ; Vol. 201, No. 2. pp. 500-505.
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