Artificial P450/reductase fusion enzymes: what can we learn from their structures?


:Cytochromes P450 constitute a superfamily of hemoproteins which have evolved from a common ancestor. Recent advances in molecular biology have offered a powerful approach to the research on the multiplicity of P450. Now, every mammalian P450 species can be characterized by heterologous expression of its cDNA. In addition, a heterogous expression system is also useful for analysis of structure-function relationships of P450 monooxygenases. Comparison of enzymatic activity and expression level in yeast of a series of artificial genetic fusion enzymes of P450 with P450 reductase has revealed the strategy of how to construct a most suitable fusion. The P450/reductase fused enzyme is a simplified monooxygenase as compared with the two enzyme systems in nature. The P450 domain of the fused enzyme is bound to the microsomes, while the reductase domain lies on the cytoplasmic side, moving flexibly. This structural feature seems to reflect the topology of both enzymes in mammalian microsomes, and will be used as a model to analyze in vivo protein-protein interactions of microsomal P450 monooxygenases. Combined with the discovery of some naturally occurring P450/reductase fusions from bacteria to mammals, comparison of these natural enzymes with artificial ones will be discussed.






Yabusaki Y




Has Abstract


1995-01-01 00:00:00














  • Use of crosslinking for revealing the DNA phosphate groups forming specific contacts with the E. coli Fpg protein.

    abstract::Specific contacts between DNA phosphate groups and positively charged nucleophilic amino acids from the Escherichia coli Fpg protein play a significant role in DNA-Fpg protein interaction. In order to identify these phosphate groups the chemical crosslinking procedure was carried out. The probing of the Fpg protein ac...


    pub_type: 杂志文章


    authors: Kuznetsova S,Rykhlevskaya A,Taranenko M,Sidorkina O,Oretskaya T,Laval J

    更新日期:2003-05-01 00:00:00

  • VaH3, one of the principal hemorrhagins in Vipera ammodytes ammodytes venom, is a homodimeric P-IIIc metalloproteinase.

    abstract::Hemorrhage is the most potent manifestation of envenomation by Vipera ammodytes ammodytes (V. a. ammodytes) venom in man. A detailed description of the venom components contributing to this effect is thus medically very important. We have characterized a novel component, termed here VaH3, as a potently hemorrhagic sna...


    pub_type: 杂志文章


    authors: Sajevic T,Leonardi A,Kovačič L,Lang-Balija M,Kurtović T,Pungerčar J,Halassy B,Trampuš-Bakija A,Križaj I

    更新日期:2013-06-01 00:00:00

  • High altitude mediated skeletal muscle atrophy: Protective role of curcumin.

    abstract::Chronic hypobaric hypoxia induced muscle atrophy results in decreased physical performance at high altitude. Curcumin has been shown to have muscle sparing effects under cachectic conditions. However, the protective effects of curcumin under chronic hypobaric hypoxia have not been studied till now. Therefore, the pres...


    pub_type: 杂志文章


    authors: Chaudhary P,Sharma YK,Sharma S,Singh SN,Suryakumar G

    更新日期:2019-01-01 00:00:00

  • RNA editing in higher plant mitochondria: analysis of biochemistry and specificity.

    abstract::RNA editing alters genomically encoded cytidines to uridines posttranscriptionally in higher plant mitochondria. Most of these editing events occur in translated regions and consequently alter the amino acid sequence. In Oenothera berteriana more than 500 editing sites have been detected and the total number of editin...


    pub_type: 杂志文章,评审


    authors: Yu W,Fester T,Bock H,Schuster W

    更新日期:1995-01-01 00:00:00

  • Positive and negative regulatory elements in the dnaA-dnaN-recF operon of Escherichia coli.

    abstract::The recF gene of E coli lies within a cluster of genes which play essential roles in DNA replication; the gene order is dnaA dnaN recF gyrB. Each of these genes has its own promoters which, with the exception of dnaA promoters, reside entirely within the translated region of the respective preceding gene. In this repo...


    pub_type: 杂志文章


    authors: Pérez-Roger I,García-Sogo M,Navarro-Aviñó JP,López-Acedo C,Macián F,Armengod ME

    更新日期:1991-02-01 00:00:00

  • Identification of multiple antimicrobial peptides from the skin of fine-spined frog, Hylarana spinulosa (Ranidae).

    abstract::In this study, peptidomics and genomics analyses were used to study antimicrobial peptides from the skin of Hylarana spinulosa. Twenty-nine different antimicrobial peptide precursors were characterized from the skin of H. spinulosa, which produce 23 mature antimicrobial peptides belonging to 12 different families. To ...


    pub_type: 杂志文章


    authors: Yang X,Hu Y,Xu S,Hu Y,Meng H,Guo C,Liu Y,Liu J,Yu Z,Wang H

    更新日期:2013-12-01 00:00:00

  • Characterization of human nicotinate phosphoribosyltransferase: Kinetic studies, structure prediction and functional analysis by site-directed mutagenesis.

    abstract::Nicotinate phosphoribosyltransferase (NaPRT, EC catalyzes the conversion of nicotinate (Na) to nicotinate mononucleotide, the first reaction of the Preiss-Handler pathway for the biosynthesis of NAD(+). Even though NaPRT activity has been described to be responsible for the ability of Na to increase NAD(+) l...


    pub_type: 杂志文章


    authors: Galassi L,Di Stefano M,Brunetti L,Orsomando G,Amici A,Ruggieri S,Magni G

    更新日期:2012-02-01 00:00:00

  • Expression of mitochondrial genes and of the transcription factors involved in the biogenesis of mitochondria Tfam, NRF-1 and NRF-2, in rat liver, testis and brain.

    abstract::Mitochondrial function requires genes encoded in both mitochondrial and nuclear genomes. Tfam, the activator of mammalian mitochondrial transcription, is encoded in the nucleus and its expression has been shown in in vitro studies to be controlled by nuclear respiratory factors NRF-1 and NRF-2. In order to understand ...


    pub_type: 杂志文章


    authors: Escrivá H,Rodríguez-Peña A,Vallejo CG

    更新日期:1999-10-01 00:00:00

  • Characterization and primary structure of proteins L28, L33 and L34 from Bacillus stearothermophilus ribosomes.

    abstract::The complete amino acid sequences of 3 proteins from the 50S subunit of Bacillus stearothermophilus ribosomes were determined by N-terminal sequence analysis and by sequencing of overlapping fragments obtained from enzymatic digestions and chemical cleavages. The proteins BstL28, BstL33 and BstL34, named according to ...


    pub_type: 杂志文章


    authors: Kruft V,Kapp U,Wittmann-Liebold B

    更新日期:1991-07-01 00:00:00

  • Regulatory properties of pigeon heart muscle AMP deaminase.

    abstract::The kinetic and regulatory properties of purified pigeon heart muscle AMP deaminase were investigated. In the presence of 100 mM potassium chloride, the enzyme exhibited a slightly sigmoidal type of kinetics. Addition of ATP to the incubation medium changed the reaction rate versus substrate concentration plot into a ...


    pub_type: 杂志文章


    authors: Kaletha K,Bogdanowicz S,Raffin JP

    更新日期:1987-02-01 00:00:00

  • Principles and equations for measuring and interpreting protein stability: From monomer to tetramer.

    abstract::The ability to measure the thermodynamic stability of proteins with precision is important for both academic and applied research. Such measurements rely on mathematical models of the protein denaturation profile, i.e. the relation between a global protein signal, corresponding to the folding states in equilibrium, an...


    pub_type: 杂志文章,评审


    authors: Bedouelle H

    更新日期:2016-02-01 00:00:00

  • Mutations in ribosomal proteins L7/L12 perturb EF-G and EF-Tu functions.

    abstract::In vitro cycling rates of E. coli ribosomes and of elongation factors EF-Tu and EF-G have been obtained and these are compatible with translation rates in vivo. We show that the rate of translocation is faster than 50 s-1 and therefore that the EF-G function is not a rate limiting step in protein synthesis. The in viv...


    pub_type: 杂志文章


    authors: Bilgin N,Kirsebom LA,Ehrenberg M,Kurland CG

    更新日期:1988-05-01 00:00:00

  • Regulation of lysophosphatidate signaling by autotaxin and lipid phosphate phosphatases with respect to tumor progression, angiogenesis, metastasis and chemo-resistance.

    abstract::Evidence from clinical, animal and cell culture studies demonstrates that increased autotaxin (ATX) expression is responsible for enhancing tumor progression, cell migration, metastases, angiogenesis and chemo-resistance. These effects depend mainly on the rapid formation of lysophosphatidate (LPA) by ATX. Circulating...


    pub_type: 杂志文章,评审


    authors: Samadi N,Bekele R,Capatos D,Venkatraman G,Sariahmetoglu M,Brindley DN

    更新日期:2011-01-01 00:00:00

  • Suppression of Neu1 sialidase delays the absorption of yolk sac in medaka (Oryzias latipes) accompanied with the accumulation of α2-3 sialo-glycoproteins.

    abstract::Sialidase catalyzes the removal of sialic acids from glycoconjugates. Recently, medaka sialidase Neu1 has been cloned and its enzymatic properties were investigated. Although enzymatic properties of this sialidase, such as optimal pH and substrate specificity, exhibits high similarity with human NEU1, Neu1 physiologic...


    pub_type: 杂志文章


    authors: Ryuzono S,Takase R,Kamada Y,Ikenaga T,Chigwechokha PK,Komatsu M,Shiozaki K

    更新日期:2017-04-01 00:00:00

  • Generation of hydrogen peroxide during the oxidation of L-phenylalanine by Proteus mirabilis isolated membranes.

    abstract::In the process of L-phenylalanine oxidation by Proteus mirabilis cytoplasmic membrane, hydrogen peroxide was produced at a rate corresponding to 1-3 per cent of the total electron flow (30-110 nmoles min-1mg-1). Peroxide was estimated using a fluorimetric assay with horseradish peroxidase, or by anodic oxidation on a ...


    pub_type: 杂志文章


    authors: Sauret-Ignazi G,Laboure-Rossat AM,Jouve HM,Pelmont J

    更新日期:1982-10-01 00:00:00

  • Stabilization of integral membrane proteins in aqueous solution using fluorinated surfactants.

    abstract::Surfactants carrying either a hydrocarbon or a fluorocarbon alkyl chain have been synthesized. The polar head was either tris(hydroxymethyl)acrylamidomethane (THAM), telomerized THAM, or a glycosylated THAM moiety. The aqueous solubility of some of these molecules was increased by oxidizing to a sulfoxide the thioethe...


    pub_type: 杂志文章


    authors: Chabaud E,Barthélémy P,Mora N,Popot JL,Pucci B

    更新日期:1998-05-01 00:00:00

  • Lipolytic activities against trioctanoin and monoolein in rat intestinal mucosa.

    abstract::The hydrolysis of trioctanoin and of monoolein by rat intestinal homogenates, catalyzed by the previously described "trioctanoin hydrolase" and "monoglyceride lipase" [Ref. 6 and 11], was shown to be entirely due to the action of the intestinal glycerol-ester hydrolase [Ref. 1 and 2] and to contaminating pancreatic li...


    pub_type: 杂志文章


    authors: Negrel R,Serrero G,Ailhaud G

    更新日期:1977-01-01 00:00:00

  • A comparative Raman spectroscopic study of cholinesterases.

    abstract::We report Raman spectra of various cholinesterases: lytic tetrameric forms (G4) obtained by tryptic digestion of asymmetric acetylcholinesterase (AChE) from Torpedo californica and Electrophorus electricus, a PI-PLC-treated dimeric form (G2) of AChE from T marmorata, and the soluble tetrameric form (G4) of butyrylchol...


    pub_type: 杂志文章


    authors: Aslanian D,Grof P,Bon S,Masson P,Négrerie M,Chatel JM,Balkanski M,Taylor P,Massoulié J

    更新日期:1991-11-01 00:00:00

  • A convenient and robust method for construction of combinatorial and random mutant libraries.

    abstract::Here we describe a convenient and robust ligase-independent method for construction of combinatorial and random mutant libraries. The homologous genes flanked by plasmid-derived DNA sequences are fragmented, and the random fragments are reassembled in a self-priming polymerase reaction to obtain chimeric genes. The pr...


    pub_type: 杂志文章


    authors: An Y,Wu W,Lv A

    更新日期:2010-08-01 00:00:00

  • A potyvirus-based gene vector allows producing active human S-COMT and animal GFP, but not human sorcin, in vector-infected plants.

    abstract::Potato virus A (PVA), a potyvirus with a (+)ssRNA genome translated to a large polyprotein, was engineered and used as a gene vector for expression of heterologous proteins in plants. Foreign genes including jellyfish GFP (Aequorea victoria) encoding the green fluorescent protein (GFP, 27 kDa) and the genes of human o...


    pub_type: 杂志文章


    authors: Kelloniemi J,Mäkinen K,Valkonen JP

    更新日期:2006-05-01 00:00:00

  • PssO, a unique extracellular protein important for exopolysaccharide synthesis in Rhizobium leguminosarum bv. trifolii.

    abstract::Synthesis and secretion of polysaccharides by Gram-negative bacteria are a result of a concerted action of enzymatic and channel-forming proteins localized in different compartments of the cell. The presented work comprises functional characterization of PssO protein encoded within the previously identified, chromosom...


    pub_type: 杂志文章


    authors: Marczak M,Mazur A,Gruszecki WI,Skorupska A

    更新日期:2008-11-01 00:00:00

  • EPR determination of the oxidation-reduction potentials of the hemes in cytochrome c3 from Desulfovibrio vulgaris.

    abstract::EPR spectroscopy in conjunction with oxidation-reduction potentiometry has been used to determine the half-reduction potentials of the four hemes of cytochrome c3. As predicted, the four hemes of cytochrome c3 have different mid-point potentials. The Em values are: Heme I,--284 mV; Heme II,--310 mV; Heme III,--324 mV ...


    pub_type: 杂志文章


    authors: Dervartanian DV,Xavier AV,Gall JL

    更新日期:1978-01-01 00:00:00

  • A new fluorometric assay for the study of DNA-binding and 3'-processing activities of retroviral integrases and its use for screening of HIV-1 integrase inhibitors.

    abstract::Fluorometry using a substrate DNA labeled with a single fluorophore (6-carboxyfluorescein) at the 3'-end of the processed strand was shown to be a useful tool for monitoring DNA-binding and 3'-processing activities of HIV-1 and PFV integrases (INs). The DNA binding to either of the INs resulted in a fluorescence signa...


    pub_type: 杂志文章


    authors: Anisenko A,Agapkina J,Zatsepin T,Yanvarev D,Gottikh M

    更新日期:2012-11-01 00:00:00

  • Biotechnology tools and applications for development of oilseed crops with healthy vegetable oils.

    abstract::Vegetable oils, consisting principally of triacylglycerols (TAG), are major sources of calories and essential fatty acids in the human diet. The fatty acid composition of TAG is a primary determinant of the nutritional quality and health-promoting properties of vegetable oils. TAG fatty acid composition also affects t...


    pub_type: 杂志文章,评审


    authors: Msanne J,Kim H,Cahoon EB

    更新日期:2020-11-01 00:00:00

  • Efficacy of azelaic acid on hepatic key enzymes of carbohydrate metabolism in high fat diet induced type 2 diabetic mice.

    abstract::Azelaic acid (AzA), a C9 linear α,ω-dicarboxylic acid, is found in whole grains namely wheat, rye, barley, oat seeds and sorghum. The study was performed to investigate whether AzA exerts beneficial effect on hepatic key enzymes of carbohydrate metabolism in high fat diet (HFD) induced type 2 diabetic C57BL/6J mice. C...


    pub_type: 杂志文章


    authors: Muthulakshmi S,Saravanan R

    更新日期:2013-06-01 00:00:00

  • Dual hydrolytic role for Pb(II) ions.

    abstract::RNA phosphodiester bonds can be cleaved by metal ions, of which Pb2+ is one of the most effective. It can cleave both generally and site-specifically, depending on the substrate and the conditions. In addition, metal ions are also known to cleave ester bonds between amino acid and the 3'-end of transfer RNA. Here we r...


    pub_type: 杂志文章


    authors: Otzen DE,Barciszewski J,Clark BF

    更新日期:1994-01-01 00:00:00

  • Molecular characterization of BjussuSP-I, a new thrombin-like enzyme with procoagulant and kallikrein-like activity isolated from Bothrops jararacussu snake venom.

    abstract::A thrombin-like enzyme, named BjussuSP-I, isolated from Bothrops jararacussu snake venom, is an acidic single-chain glycoprotein with M(r)=61,000, pI approximately 3.8 and 6% sugar. BjussuSP-I shows high proteolytic activity upon synthetic substrates, such as S-2238 and S-2288. It also shows procoagulant and kallikrei...


    pub_type: 杂志文章


    authors: Sant'Ana CD,Bernardes CP,Izidoro LF,Mazzi MV,Soares SG,Fuly AL,Zingali RB,Magro AJ,Braz AS,Fontes MR,Stábeli RG,Sampaio SV,Soares AM

    更新日期:2008-03-01 00:00:00

  • Telomere dynamics in human cells.

    abstract::Human telomeres are intrinsically dynamic structures, with multiple biological processes operating to generate substantial length heterogeneity. Processes that operate specifically at the terminus, that include the end-replication problem coupled with C-strand resection, result in gradual telomere erosion with ongoing...


    pub_type: 杂志文章,评审


    authors: Baird DM

    更新日期:2008-01-01 00:00:00

  • Control, regulation and thermodynamics of free-energy transduction.

    abstract::The quantitative formalism called Metabolic Control Theory makes it possible to be precise in discussions of metabolic control. To illustrate this, I will mention 2 experimental systems where free energy is converted from one form to another, i.e., bacteriorhodopsin liposomes and mitochondrial oxidative phosphorylatio...


    pub_type: 杂志文章,评审


    authors: Westerhoff HV

    更新日期:1989-08-01 00:00:00

  • The biosynthesis of highly branched N-glycans: studies on the sequential pathway and functional role of N-acetylglucosaminyltransferases I, II, III, IV, V and VI.

    abstract::At least 6 N-acetylglucosaminyltransferases (GlcNAc-T I, II, III, IV, V and VI) are involved in initiating the synthesis of the various branches found in complex asparagine-linked oligosaccharides (N-glycans), as indicated below: GlcNAc beta 1-6 GlcNAc-T V GlcNAc beta 1-4 GlcNAc-T VI GlcNAc beta 1-2Man alpha 1-6 GlcNA...


    pub_type: 杂志文章


    authors: Brockhausen I,Narasimhan S,Schachter H

    更新日期:1988-11-01 00:00:00