1
40
1
-
Text
A resource consisting primarily of words for reading. Examples include books, letters, dissertations, poems, newspapers, articles, archives of mailing lists. Note that facsimiles or images of texts are still of the genre Text.
URL Address
n/a
Rights
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
Pages
399-405
Issue
4
Volume
25
Search for Full-text
Locate full-text within NEOMED Library's e-journal collections
<p>Users with a NEOMED Library login can search for full-text journal articles at the following url: <a href="https://libraryguides.neomed.edu/home">https://libraryguides.neomed.edu/home</a></p>
Dublin Core
The Dublin Core metadata element set is common to all Omeka records, including items, files, and collections. For more information see, http://dublincore.org/documents/dces/.
Title
A name given to the resource
Coexpression of cytochrome P4502A6 and human NADPH-P450 oxidoreductase in the baculovirus system
Publisher
An entity responsible for making the resource available
Drug Metabolism and Disposition
Date
A point or period of time associated with an event in the lifecycle of the resource
1997
1997-04
Subject
The topic of the resource
human; Pharmacology & Pharmacy; protein; enzymes; reductase; liver-microsomes; catalytic properties; cdna-directed expression; insect cells; nuclear polyhedrosis-virus; promoters; vector system
Creator
An entity primarily responsible for making the resource
Chen L P; Buters J T M; Hardwick J P; Tamura S; Penman B W; Gonzalez F J; Crespi C L
Description
An account of the resource
Heterologous expression using baculovirus vectors has become a popular method for the production of catalytically active cytochrome P450s (CYPs). We have systematically optimized the multiplicity of infection (MOI) for a coinfection approach for the coexpression of CYP2A6 (viral vector designated v2A6) and NADPH-P450 oxidoreductase (OR; viral vector designated vOR) using Sf9 insect cells. A 3000-fold range of MOI was examined in stationary culture and stirred suspension culture. Surprisingly, our results Indicate that the best CYP2A6 catalytic activity (850-1300 pmol/ min/mg total lysate protein as measured by coumarin 7-hydroxylase activity) was obtained only when using a low MOI of v2A6 (1.5-3 x 10(-2)) and a vOR of 10- to 20-fold less, This activity was similar to 7- to 11-fold higher than the best activity obtained when infecting cells with v2A6 alone. At this level of coinfection, the P450 content ranged from 180 to 250 pmol/mg total lysate protein, and the NADPH cytochrome c reductase activity ranged from 350 to 520 nmol/min/mg total lysate protein. Increasing the MOI of both viruses to 50-fold higher resulted in lower overall activity with the optimum (250 pmol/min/mg total lysate protein) being seen earlier postinfection (60 vs. 72 hr). Increasing the MOI of vOR to levels comparable with those of v2A6, decreased coumarin 7-hydroxylase activity 14-fold. These results suggest that the best CYP2A6 catalytic activity depends on properly posttranslationally modified proteins accumulating in a right ratio as a result of primary, secondary, and possibly tertiary infection of both viruses. These results also suggest that high OR expression results in degradation of P450.
Identifier
An unambiguous reference to the resource within a given context
n/a
Format
The file format, physical medium, or dimensions of the resource
Journal Article or Conference Abstract Publication
1997
Buters J T M
catalytic properties
cdna-directed expression
Chen L P
Crespi C L
Drug Metabolism and Disposition
Enzymes
Gonzalez F J
Hardwick J P
Human
insect cells
Journal Article or Conference Abstract Publication
liver-microsomes
nuclear polyhedrosis-virus
Penman B W
Pharmacology & Pharmacy
promoters
Protein
reductase
Tamura S
vector system