Rabbit P450 2E1 Expressed in CHO-K1 Cells Has a Short Half-Life

S. Barmada, E. Kienle, Dennis Koop

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Rabbit P450 2E1 was stably expressed in Chinese hamster ovary cells after cotransfection with pRC/CMV-2E1 and pFR400 which expresses murine dihydrofolate reductase with a single arginine to leucine substitution at position 22. This mutation permits amplification of expression with increasing methotrexate concentrations in CHO-K1 cells that are not dihydrofolate reductase deficient. After amplification with 1 μM methotrexate, a representative clone expressed about 15 pmol of P450 2E1/mg microsomal protein. Cells from a single 35-mm plate catalyzed the formation of 1.02 nmol 6-hydroxychlorzoxazone/106 cells/h or about 127 pmol/mg total cell protein/min. The enzyme was rapidly labeled when pulsed with [35S]-methionine. Initial pulse-chase experiments indicate that the expressed protein has a half-life of 4.8 h.

Original languageEnglish (US)
Pages (from-to)601-607
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume206
Issue number2
DOIs
StatePublished - Jan 17 1995

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CHO Cells
Half-Life
Tetrahydrofolate Dehydrogenase
Rabbits
Methotrexate
Amplification
Proteins
Leucine
Methionine
Arginine
Substitution reactions
Cricetulus
Cells
Ovary
Clone Cells
Enzymes
Mutation
Experiments

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology
  • Biophysics
  • Biochemistry

Cite this

Rabbit P450 2E1 Expressed in CHO-K1 Cells Has a Short Half-Life. / Barmada, S.; Kienle, E.; Koop, Dennis.

In: Biochemical and Biophysical Research Communications, Vol. 206, No. 2, 17.01.1995, p. 601-607.

Research output: Contribution to journalArticle

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