Characterization and purification of a lipoxygenase inhibitor in human epidermoid carcinoma A431 cells

C. J. Chen, H. S. Huang, Y. T. Lee, C. Y. Yang, W. C. Chang

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

A lipoxygenase inhibitor in the cytosolic fraction of human epidermoid carcinoma A431 cells was characterized and purified. The cytosolic inhibitor lost the inhibitory activity upon heating at 75°C for 15 min or pretreating with 1 mg/ml trypsin at 37°C for 60 min. Cytosol, after dialysis, lost the inhibitory activity but its inhibitory activity recovered when 1 mM GSH was added to the dialysate. The inhibitory activity of cytosol was also abolished by treatment either with 1 mM iodoacetate at 4°C for 1 h or with 0.5 mM H2O2. The pI of the inhibitor was approx. 7.0. In addition to 12-lipoxygenase, the inhibitor inhibited the activities of 5-lipoxygenase and fatty acid cyclo-oxygenase in a cell-free system. The inhibitor was purified by a series of column chromatographies using CM Sephadex C-50, Sephadex G-100 SF and Mono P columns. A major 22 kDa protein was obtained that was distinct from selenium-dependent glutathione peroxidase.

Original languageEnglish
Pages (from-to)193-198
Number of pages6
JournalBiochemical Journal
Volume327
Issue number1
Publication statusPublished - 1997
Externally publishedYes

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Lipoxygenase Inhibitors
Cytosol
Purification
Squamous Cell Carcinoma
Iodoacetates
Arachidonate 5-Lipoxygenase
Column chromatography
Dialysis
Cell-Free System
Dialysis Solutions
Prostaglandin-Endoperoxide Synthases
Selenium
Glutathione Peroxidase
Heating
Trypsin
Chromatography
Proteins
sephadex

ASJC Scopus subject areas

  • Biochemistry

Cite this

Characterization and purification of a lipoxygenase inhibitor in human epidermoid carcinoma A431 cells. / Chen, C. J.; Huang, H. S.; Lee, Y. T.; Yang, C. Y.; Chang, W. C.

In: Biochemical Journal, Vol. 327, No. 1, 1997, p. 193-198.

Research output: Contribution to journalArticle

Chen, C. J. ; Huang, H. S. ; Lee, Y. T. ; Yang, C. Y. ; Chang, W. C. / Characterization and purification of a lipoxygenase inhibitor in human epidermoid carcinoma A431 cells. In: Biochemical Journal. 1997 ; Vol. 327, No. 1. pp. 193-198.
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