Relationship between cell surface protease activity and doubling time in various normal and transformed cells

Victor Bernard Hatcher, Michael S. Wertheim, Choo Y. Rhee, Grace Tsien, Peter G. Burk

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

A sensitive method for measuring cell surface and secreted protease activity utilizing 3H-labelled casein is described. The method is based upon proteolytic degradation of the casein substrate into trichloroacetic acid soluble 3H-labelled peptides. Utilizing the radioassay we found that all cultured cell lines examined contain cell surface proteolytic activity which is not secreted into the media. The protease activity was found to be due to protease(s) other than plasminogen activator or plasmin. A comparison of surface protease activity of normal and transformed mouse epidermal cells indicated that the transformed cells contained approximately 3-4 times more proteolytic activity than the normal cells. Surface protease activity was also correlated with the doubling times of various cultured cells. The results indicated that cultured cells with doubling times of greater than three days possess less surface protease activity than cells with shorter doubling times. In order to determine changes in the levels of surface protease activity during the cell cycle several cell lines were synchronized. In synchronized rabbit aortic fibroblasts, mouse transformed epidermal cells and human melanoma cells, a marked increase in surface protease activity was observed during or before mitosis. The protease levels decreased following mitosis. The results suggest that in culture, cell surface protease(s) may be important factor in regulating the rate of cell growth.

Original languageEnglish (US)
Pages (from-to)499-510
Number of pages12
JournalBBA - General Subjects
Volume451
Issue number2
DOIs
StatePublished - Dec 21 1976

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Peptide Hydrolases
Cells
Cultured Cells
Caseins
Mitosis
Cell Line
Trichloroacetic Acid
Plasminogen Activators
Fibrinolysin
Cell growth
Fibroblasts
Cell culture
Melanoma
Cell Cycle
Cell Culture Techniques
Rabbits
Degradation
Peptides
Substrates
Growth

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Medicine(all)

Cite this

Relationship between cell surface protease activity and doubling time in various normal and transformed cells. / Hatcher, Victor Bernard; Wertheim, Michael S.; Rhee, Choo Y.; Tsien, Grace; Burk, Peter G.

In: BBA - General Subjects, Vol. 451, No. 2, 21.12.1976, p. 499-510.

Research output: Contribution to journalArticle

Hatcher, Victor Bernard ; Wertheim, Michael S. ; Rhee, Choo Y. ; Tsien, Grace ; Burk, Peter G. / Relationship between cell surface protease activity and doubling time in various normal and transformed cells. In: BBA - General Subjects. 1976 ; Vol. 451, No. 2. pp. 499-510.
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