High Conformational Stability of Cytochrome P-450 1A2. Evidence from UV Absorption Spectra

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Title:High Conformational Stability of Cytochrome P-450 1A2. Evidence from UV Absorption Spectra
Creators:
Anzenbacher, Pavel
Bec, Nicole
Hudeček, Jiří
Lange, Reinhard
Anzenbacherová, Eva
Journal or Publication Title:
Collection of Czechoslovak Chemical Communications, 63, 3, pp. 441-448
Uncontrolled Keywords:Conformation, α-Naphthoflavone, CYP1A2, Derivative spectra, Cytochrome P-450

Abstract

The fourth derivative of absorption spectra between 260 and 310 nm were used for monitoring the changes in exposure of tyrosine and tryptophan side chains in cytochrome P-450 1A2 to solvent. Titration of the enzyme with a specific inhibitor, α-naphthoflavone (2-phenylazo[<i>h</i>]chromen-4-one) to inhibitor concentration of 30 μM resulted in small but pronounced changes in derivative spectra (decrease in the maximum amplitude, downshift of the spectral maximum at about 293 nm) corresponding to the exposure of tryptophans towards the solvent. Further addition of the inhibitor led to a decrease of the exposure of these aromatic side-chains. Similar behaviour was also observed in this work for other cytochromes P-450 (2B4 and 11A1). The fourth derivative of absorption spectra was also used to examine the stability of the enzyme both in the presence and absence of α-naphthoflavone, with increasing pressure (up to 400 MPa) and temperature (up to 35 °C) as pertubing factors. The results show that cytochrome P-450 1A2 has a stable conformation as all the conformational changes observed were (spectrally) fully reversible.

Title:High Conformational Stability of Cytochrome P-450 1A2. Evidence from UV Absorption Spectra
Creators:
Anzenbacher, Pavel
Bec, Nicole
Hudeček, Jiří
Lange, Reinhard
Anzenbacherová, Eva
Uncontrolled Keywords:Conformation, α-Naphthoflavone, CYP1A2, Derivative spectra, Cytochrome P-450
Divisions:Life and Chemical Sciences > Institute of Organic Chemistry and Biochemistry > Collection of Czechoslovak Chemical Communications
Journal or Publication Title:Collection of Czechoslovak Chemical Communications
Volume:63
Number:3
Page Range:pp. 441-448
ISSN:0010-0765
E-ISSN:1212-6950
Publisher:Institute of Organic Chemistry and Biochemistry
Related URLs:
URLURL Type
http://dx.doi.org/10.1135/cccc19980441UNSPECIFIED
ID Code:1312
Item Type:Article
Deposited On:06 Feb 2009 17:08
Last Modified:06 Feb 2009 16:08

Citation

Anzenbacher, Pavel; Bec, Nicole; Hudeček, Jiří; Lange, Reinhard; Anzenbacherová, Eva (1998) High Conformational Stability of Cytochrome P-450 1A2. Evidence from UV Absorption Spectra. Collection of Czechoslovak Chemical Communications, 63 (3). pp. 441-448. ISSN 0010-0765

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