Regular ArticleA Truncation of 2B Subfamily Cytochromes P450 Yields Increased Expression Levels, Increased Solubility, and Decreased Aggregation While Retaining Function☆
References (46)
- et al.
Biochem. Biophys. Res. Commun.
(2001) - et al.
Arch. Bioch. Biophys.
(1996) - et al.
Mol. Cell
(2000) - et al.
J. Biol. Chem.
(1991) - et al.
J. Biol. Chem.
(1993) - et al.
Arch. Biochem. Biophys.
(1993) - et al.
Arch. Biochem. Biophys.
(1995) - et al.
Arch. Biochem. Biophys.
(1997) - et al.
J. Biol. Chem.
(2000) - et al.
J. Biol. Chem.
(1997)
Arch. Biochem. Biophys.
Arch. Biochem. Biophys.
Arch. Biochem. Biophys.
Arch. Biochem. Biophys.
Arch. Biochem. Biophys.
J. Biol. Chem.
Arch. Biochem. Biophys.
Arch. Biochem. Biophys.
Biochem. Biophys. Res. Commun.
Curr. Drug Metab.
Drug Metab. Rev.
Drug Metab. Dispos.
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2016, Biophysical ChemistryCitation Excerpt :This is also the background in which all mutations were made. CYP2B4 enzymes were expressed in Escherichia coli TOPP3 cells as previously described [13]. Protein expression was induced by adding IPTG, 0.5 mM) and ALA (1 mM) to Terrific broth medium (A600 ~ 0.7 at 37 °C) in the presence of ampicillin (100 mg/ml).
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This work was supported by a National Research Service Award GM20674 (E.E.S.), ES03619 (J.R.H.), and Center Grant ES06676.
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