Document Detail


Baculovirus-directed expression of rabbit UDP-glucuronosyltransferases in Spodoptera frugiperda cells.
MedLine Citation:
PMID:  9193877     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The rabbit liver UDP-glucuronosyltransferase (UGT) cDNAs that encode the 4-hydroxybiphenyl UGT2B13 and 4-nitrophenol UGT1A6 have been cloned into baculovirus. Spodoptera frugiperda (SF-9) cells infected with the UGT recombinant baculovirus produced significant amounts of protein, which was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblot analysis, and pulse chase with 35S-amino acids. The expression of the UGT proteins in SF-9 cells were detected at approximately 24 hr postinfection, with maximal levels of protein seen at 48 hr. Immunoprecipitation of newly synthesized 35S-labeled proteins demonstrated that the maximal rate of protein synthesis in SF-9 cells infected with the UGT baculovirus occur at 48 hr postinfection, although the proteins are abundant in the cells for up to 96 hr. When compared with the expression levels of the same cDNAs through transient transfection into COS-1 cells, the insect-derived UGT proteins showed nearly 50- to 100-fold greater protein accumulation. Although kinetic analysis demonstrated that turnover rate of the SF-9-expressed proteins were greater than their counterparts in COS-1 cells, K(M) values for UGT1A6 and UGT2B13 in SF-9 and COS-1 cells were similar. Overall, SF-9 cells seem to serve as an efficient expression system for the production of the mammalian UGTs.
Authors:
N Nguyen; R H Tukey
Related Documents :
6154527 - Inhibition by 2',3'-dideoxythymidine of retroviral infection of mouse and human cells.
11412917 - Use of percoll for the infection of cells in vitro with cryptosporidium parvum oocysts.
11090177 - Striking similarity of murine nectin-1alpha to human nectin-1alpha (hvec) in sequence a...
6093367 - Characterisation of a herpes simplex virus type 1 mutant which has a temperature-sensit...
23552737 - Selective growth inhibition by glycogen synthase kinase-3 inhibitors in tumorigenic hel...
19148897 - Metabolism of anandamide by cox-2 is necessary for endocannabinoid-induced cell death i...
Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Drug metabolism and disposition: the biological fate of chemicals     Volume:  25     ISSN:  0090-9556     ISO Abbreviation:  Drug Metab. Dispos.     Publication Date:  1997 Jun 
Date Detail:
Created Date:  1997-08-15     Completed Date:  1997-08-15     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  9421550     Medline TA:  Drug Metab Dispos     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  745-9     Citation Subset:  IM    
Affiliation:
Department of Pharmacology, University of California-San Diego, Cancer Center, La Jolla 92093-0636, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Animals
Baculoviridae / genetics*
Blotting, Western
COS Cells
Cells, Cultured
DNA, Complementary
Gene Expression Regulation, Viral*
Glucuronosyltransferase / genetics*,  metabolism
Rabbits
Spodoptera / virology
Time Factors
Grant Support
ID/Acronym/Agency:
GM49135/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/DNA, Complementary; EC 2.4.1.17/Glucuronosyltransferase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Hydroxylation and demethylation of the tricyclic antidepressant nortriptyline by cDNA-expressed huma...
Next Document:  In vivo induction and in vitro inhibition of hepatic cytochrome P450 activity by the benzodiazepine ...