FTIR studies of the redox partner interaction in cytochrome P450: The Pdx–P450cam couple

Karyakin, Andrey; Motiejunas, Domantas; Wade, Rebecca C.; Jung, Christiane

doi:10.1016/j.bbagen.2006.08.020

Cited by 16 publications

(18 citation statements)

References 58 publications

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“…Applying the above-explained procedure, we reported the first ab initio quantum chemistry description of the electron migration between the two active sites [85]. Starting from the two separate crystal structures of P450cam and Pdx, we used the docking program pyDock [86] as well as the protein modeling program PELE [13] to generate a few complex structures having iron-iron distances between 12.7 and 15.7 Å and being very similar to the structures proposed in the literature [87][88][89]. The results of mapping the excess electron transfer pathway between the Fe 2 S 2 cluster of Pdx and the heme of P450cam clearly identify residue Asp38 of Pdx and Arg112 of P450cam as the key residues within the electron transfer pathway.…”

Section: P450cam-pdx Electron Transfermentioning

confidence: 96%

QM/MM methods: Looking inside heme proteins biochemisty

Guallar

Wallrapp

2010

Biophysical Chemistry

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Section: P450cam-pdx Electron Transfermentioning

confidence: 96%

QM/MM methods: Looking inside heme proteins biochemisty

Guallar

Wallrapp

2010

Biophysical Chemistry

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“…Structural models of PdR-Pdx and Pdx-CYP101A1 complexes have been proposed from the structures of the individual components and spectro-scopic data (52,65,69,70). The structures of cross-linked AdR and Adx, and a related ONFR-ferredoxin (BphA4-BphA3) complex from the biphenyl dioxygenase system of Acidocorax sp.…”

Section: Discussionmentioning

confidence: 99%

“…The Arx surface potential in the vicinity of the [2Fe-2S] cluster is mainly negative, with a neutral area immediately surrounding the cluster binding loop ( Extensive studies suggested that the interaction between CYP101A1 and Pdx is dominated by contacts between Trp 106 (C-terminal residue) and Asp 38 (before the cluster binding loop) on Pdx with residues on CYP101A1 (65,69,70 Fig. S14c) (27).…”

mentioning

confidence: 99%

Molecular Characterization of a Class I P450 Electron Transfer System from Novosphingobium aromaticivorans DSM12444

Yang

Bell

Wang

et al. 2010

Journal of Biological Chemistry

135

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Cytochrome P450 (CYP) 4 enzymes constitute a superfamily of heme-containing monooxygenases (1, 2) that participate in a variety of biological processes such as carbon source assimilation, biosynthesis and biodegradation, xenobiotic detoxification, and metabolism of medicines (1, 2). The most common activity of P450 enzymes is the insertion of an oxygen atom from dioxygen into chemically inert carbon-hydrogen bonds, but other reaction types including dealkylation, desaturation, heteroatom oxidation, epoxidation, phenol coupling, and reductive dehalogenation are also known (3-6). The various activities of P450 enzymes are of great interest due to their potential applications in, for example, synthesis of fine chemicals and drug metabolites under mild conditions with high specificity.P450 enzymatic activity requires two electrons that are usually derived from NAD(P)H and delivered to the P450s by electron transfer proteins which are broadly divided into two classes (7,8). Class I systems are diverse, usually consisting of an oxygenase-coupled NAD(P)H-dependent ferredoxin reductase (ONFR) and an iron-sulfur ferredoxin. Such systems are the predominant forms in prokaryotes but are also found in eukaryotic mitochondrial membranes. ONFRs typically contain an FAD cofactor. Ferredoxin cluster types include [2Fe-2S], [3Fe-4S], [4Fe-4S], and combinations of these (7, 9). Non-ferredoxin FMN proteins have also been identified (10). Class II P450 enzymes are most common in eukaryotes and utilize an NADPH-cytochrome P450 reductase (CPR) containing prosthetic groups FAD and FMN (11). Recently other more diverse electron transfer systems for P450 enzymes have been discovered and these have been defined into several new classes (7,8).An important difference between the two main classes is that, whereas a single CPR supports the activity of all 57 human P450s and yeast CPRs support the activity of numerous P450s heterologously expressed in the organism, most class I systems show redox partner specificity. Putidaredoxin (Pdx) is well known to have an effector role in CYP101A1 activity (12, 13). The activity of CYP199A2 from Rhodopseudomonas palustris CGA009 has been reconstituted with palustrisredoxin (Pux), a [2Fe-2S] ferredoxin genomically associated with CYP199A2, and an ONFR, palustrisredoxin reductase (PuR) (14). The high demethylation activity of this system is severely compromised in the hybrid PdR/Pdx/CYP199A2 system (14) due to weak ferredoxin/P450 binding (14,15). Numerous P450 enzymes with potentially interesting and desirable activities are

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“…The Amide I band of a protein depends on the tertiary structure of the protein, with distinct m(C=O) mode frequencies for ahelices, turns, or b-sheet structures (Krimm and Bandekar 1986; Venyaminov and Kalnin 1990b), corresponding to different hydrogen bonding patterns and electrostatic interactions of the peptide carbonyl groups in these structures. The structural analysis of proteins thus largely involves IR spectroscopy, notably to inspect the effect of temperature on protein folding (Hauser et al 2008;Mukherjee et al 2008) or the consequences of protein-protein interactions (Karyakin et al 2007).…”

Section: Absorption Of Proteins In Solutionmentioning

confidence: 99%

Fourier transform infrared (FTIR) spectroscopy

2009

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Fourier transform infrared (FTIR) spectroscopy probes the vibrational properties of amino acids and cofactors, which are sensitive to minute structural changes. The lack of specificity of this technique, on the one hand, permits us to probe directly the vibrational properties of almost all the cofactors, amino acid side chains, and of water molecules. On the other hand, we can use reaction-induced FTIR difference spectroscopy to select vibrations corresponding to single chemical groups involved in a specific reaction. Various strategies are used to identify the IR signatures of each residue of interest in the resulting reaction-induced FTIR difference spectra. (Specific) Isotope labeling, site-directed mutagenesis, hydrogen/deuterium exchange are often used to identify the chemical groups. Studies on model compounds and the increasing use of theoretical chemistry for normal modes calculations allow us to interpret the IR frequencies in terms of specific structural characteristics of the chemical group or molecule of interest. This review presents basics of FTIR spectroscopy technique and provides specific important structural and functional information obtained from the analysis of the data from the photosystems, using this method.

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FTIR studies of the redox partner interaction in cytochrome P450: The Pdx–P450cam couple

Cited by 16 publications

References 58 publications

QM/MM methods: Looking inside heme proteins biochemisty

QM/MM methods: Looking inside heme proteins biochemisty

Molecular Characterization of a Class I P450 Electron Transfer System from Novosphingobium aromaticivorans DSM12444

Fourier transform infrared (FTIR) spectroscopy

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