Regulated expression of cytochrome P-450scc (cholesterol-side-chain cleavage enzyme) in cultured cell lines detected by antibody against bacterially expressed human protein

M. C. Hu, I. C. Guo, J. H. Lin, Bon Chu Chung

Research output: Contribution to journalArticle

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Abstract

The first step in the synthesis of steroids is catalysed by cytochrome P-450ssc (cholesterol-side-chain cleavage enzyme). We have investigated the synthesis of this enzyme in three cultured cell lines at the protein and hormone secretion levels. Hormone levels were measured by an enzyme immnnoassay using a monoclonal antibody against progesterone. The protein level was detected using polyclonal antibodies directed against a P-450scc fusion protein overproduced in Escherichia coli. Utilizing a bacteriophage T7 promoter expression system, a large amount of human P-450scc fusion protein was produced and easily purified. P-450scc was synthesized in the mouse adrenal tumour cell line Y1 and human choriocarcinoma cell line JEG-3, but not in monkey kidney cell line COS-1. The production of P-450scc in Y1 and JEG-3 cells was stimulated by 8-bromo cyclic AMP, the effect of which was not observed until 6 h after induction and was more pronounced at 24 h. Y1 and JEG-3 cells exhibited a difference in progesterone secretion after induction.

Original languageEnglish
Pages (from-to)813-817
Number of pages5
JournalBiochemical Journal
Volume274
Issue number3
DOIs
Publication statusPublished - Jan 1 1991

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Cholesterol Side-Chain Cleavage Enzyme
Cytochromes
Cultured Cells
Cells
Cell Line
Antibodies
Progesterone
Proteins
Fusion reactions
Hormones
Bacteriophage T7
8-Bromo Cyclic Adenosine Monophosphate
Choriocarcinoma
Bacteriophages
Glandular and Epithelial Neoplasms
Enzymes
Tumor Cell Line
Escherichia coli
Haplorhini
Tumors

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Regulated expression of cytochrome P-450scc (cholesterol-side-chain cleavage enzyme) in cultured cell lines detected by antibody against bacterially expressed human protein. / Hu, M. C.; Guo, I. C.; Lin, J. H.; Chung, Bon Chu.

In: Biochemical Journal, Vol. 274, No. 3, 01.01.1991, p. 813-817.

Research output: Contribution to journalArticle

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