|
rdf:type |
|
|
lifeskim:mentions |
|
|
pubmed:issue |
2
|
|
pubmed:dateCreated |
1995-2-16
|
|
pubmed:abstractText |
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 microM 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/10(6) 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.
|
|
pubmed:grant |
|
|
pubmed:language |
eng
|
|
pubmed:journal |
|
|
pubmed:citationSubset |
IM
|
|
pubmed:chemical |
|
|
pubmed:status |
MEDLINE
|
|
pubmed:month |
Jan
|
|
pubmed:issn |
0006-291X
|
|
pubmed:author |
|
|
pubmed:issnType |
Print
|
|
pubmed:day |
17
|
|
pubmed:volume |
206
|
|
pubmed:owner |
NLM
|
|
pubmed:authorsComplete |
Y
|
|
pubmed:pagination |
601-7
|
|
pubmed:dateRevised |
2007-11-14
|
|
pubmed:meshHeading |
pubmed-meshheading:7826377-Animals,
pubmed-meshheading:7826377-Blotting, Northern,
pubmed-meshheading:7826377-CHO Cells,
pubmed-meshheading:7826377-Cricetinae,
pubmed-meshheading:7826377-Cytochrome P-450 CYP2E1,
pubmed-meshheading:7826377-Cytochrome P-450 Enzyme System,
pubmed-meshheading:7826377-Half-Life,
pubmed-meshheading:7826377-Kinetics,
pubmed-meshheading:7826377-Methotrexate,
pubmed-meshheading:7826377-Mice,
pubmed-meshheading:7826377-Mutagenesis, Site-Directed,
pubmed-meshheading:7826377-Oxidoreductases, N-Demethylating,
pubmed-meshheading:7826377-Plasmids,
pubmed-meshheading:7826377-Point Mutation,
pubmed-meshheading:7826377-RNA, Messenger,
pubmed-meshheading:7826377-Rabbits,
pubmed-meshheading:7826377-Recombinant Proteins,
pubmed-meshheading:7826377-Tetrahydrofolate Dehydrogenase,
pubmed-meshheading:7826377-Transfection
|
|
pubmed:year |
1995
|
|
pubmed:articleTitle |
Rabbit P450 2E1 expressed in CHO-K1 cells has a short half-life.
|
|
pubmed:affiliation |
Department of Pharmacology, Oregon Health Sciences University, Portland 97206.
|
|
pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
|
