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1.
Sakamoto T Kawabe A Tokida-Segawa A Shimizu B Takatsuto S Shimada Y Fujioka S Mizutani M 《The Plant journal : for cell and molecular biology》2011,67(1):1-12
Catabolism of brassinosteroids regulates the endogenous level of bioactive brassinosteroids. In Arabidopsis thaliana, bioactive brassinosteroids such as castasterone (CS) and brassinolide (BL) are inactivated mainly by two cytochrome P450 monooxygenases, CYP734A1/BAS1 and CYP72C1/SOB7/CHI2/SHK1; CYP734A1/BAS1 inactivates CS and BL by means of C-26 hydroxylation. Here, we characterized CYP734A orthologs from Oryza sativa (rice). Overexpression of rice CYP734As in transgenic rice gave typical brassinosteroid-deficient phenotypes. These transformants were deficient in both the bioactive CS and its precursors downstream of the C-22 hydroxylation step. Consistent with this result, recombinant rice CYP734As utilized a range of C-22 hydroxylated brassinosteroid intermediates as substrates. In addition, rice CYP734As can catalyze hydroxylation and the second and third oxidations to produce aldehyde and carboxylate groups at C-26 in vitro. These results indicate that rice CYP734As are multifunctional, multisubstrate enzymes that control the endogenous bioactive brassinosteroid content both by direct inactivation of CS and by the suppression of CS biosynthesis by decreasing the levels of brassinosteroid precursors. 相似文献
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Dueckershoff K Unger M Frank A Gillam EM Guengerich FP Warzecha H 《FEBS letters》2005,579(11):2480-2484
In this study, the human cytochrome P450 (CYP) 2A6 was used in order to modify the alkaloid production of tobacco plants. The cDNA for human CYP2A6 was placed under the control of the constitutive 35S promoter and transferred into Nicotiana tabacum via Agrobacterium-mediated transformation. Transgenic plants showed formation of the recombinant CYP2A6 enzyme but no obvious phenotypic changes. Unlike wild-type tobacco, the transgenic plants accumulated cotinine, a metabolite which is usually formed from nicotine in humans. This result substantiates that metabolic engineering of the plant secondary metabolism via mammalian P450 enzymes is possible in vivo. 相似文献
4.
Inge N.A. Van Bogaert Marjan Demey Dirk Develter Wim Soetaert & Erick J. Vandamme 《FEMS yeast research》2009,9(1):87-94
Three cytochrome P450 monooxygenases belonging to the CYP52 family were isolated from the genome of the sophorolipid-producing yeast Candida bombicola using degenerate PCR and genomic walking. One gene displayed high identity with the CYP52E members and was classified into this group ( CYP52E3 ), whereas the other genes belonged to new groups: CYP52M and CYP52N. CYP52E3 and CYP52N1 turned out to be of no relevance for sophorolipid production, but show clear upregulation when the yeast cells are grown on alkanes as the sole carbon source. On the other hand, CYP52M1 is clearly upregulated during sophorolipid synthesis and very likely takes part in sophorolipid formation. 相似文献
5.
烟草细胞色素P450的基因组学分析 总被引:1,自引:0,他引:1
细胞色素P450是一类含血红素的单加氧酶超基因家族, 在植物多种代谢途径中起着重要作用。为了解烟草中的P450的种类和数量, 文章将植物代表性P450蛋白质序列与烟草基因组序列比对, 在烟草基因组中鉴定了44个P450家族共263个成员。将这些烟草P450基因与烟草表达序列标签(EST)比对, 发现173个成员有EST证据。通过与拟南芥中已知的P450蛋白序列比较, 分析了部分烟草P450蛋白序列的特征和二级结构。根据烟草基因芯片数据和部分基因的RT-PCR结果, 发现73个烟草P450基因能够在不同的生长发育时期表达, 其中部分基因具有组织特异性。这些研究结果为烟草P450基因功能的深入分析奠定了基础。 相似文献
6.
X‐ray crystal structure of cytochrome P450 monooxygenase CYP101J2 from Sphingobium yanoikuyae strain B2
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The cytochrome P450 monooxygenases (P450s) catalyze a vast array of oxygenation reactions that can be useful in biocatalytic applications. CYP101J2 from Sphingobium yanoikuyae is a P450 that catalyzes the hydroxylation of 1,8‐cineole. Here we report the crystallization and X‐ray structure elucidation of recombinant CYP101J2 to 1.8 Å resolution. The CYP101J2 structure shows the canonical P450‐fold and has an open conformation in the absence of substrate. Analysis of the structure revealed that CYP101J2, in the absence of substrate, forms a well‐ordered substrate‐binding channel that suggests a unique form of substrate guidance in comparison to other bacterial 1,8‐cineole‐hydroxylating P450 enzymes. Proteins 2017; 85:945–950. © 2016 Wiley Periodicals, Inc. 相似文献
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Yu. D. Ivanov A. V. Ivanov A. L. Kaysheva V. G. Zgoda S. A. Usanov G. Hui-Bon-Hoa A. I. Archakov 《Biochemistry (Moscow) Supplemental Series B: Biomedical Chemistry》2009,3(2):183-197
The equilibrium dissociation constants KD, the complex association / dissociation rate constants (k on /k off) and lifetimes of the complexes of redox partners were measured for three cytochrome P450-containing monooxygenase systems (P450cam, P450scc, and P450 2B4) under hydroxylation conditions. The Q parameter representing the ratio of protein-protein complex lifetime (τ lT ) to time required for a single hydroxylation cycle (τturnover) was introduced for estimation of productivity of complexes formed within the systems studied. The Q parameter was insignificantly changed upon transition from the oxidation to hydroxylation conditions. Lifetimes (τ lT ) for the binary complexes formed within the P450cam and the P450scc systems obligatory requiring an intermediate electron transfer protein between the reductase and cytochrome P450 could not realize hydroxylation reactions for substrates with known τturnover and so they were non-productive while the binary complexes formed within the P450 2B4 system, not requiring such intermediate electron-transfer protein, appeared to be productive. Formation of ternary complexes was demonstrated under hydroxylation conditions in all three systems. Analysis of Q values led to the conclusion that the ternary complexes formed within the P450cam and the P450scc systems were productive. In the case of the P450 2B4 system, more than half (about 60%) ternary complexes were also found to be productive. 相似文献
8.
Rupasinghe S Schuler MA Kagawa N Yuan H Lei L Zhao B Kelly SL Waterman MR Lamb DC 《FEBS letters》2006,580(27):6338-6342
In this work, we have spectroscopically characterised CYP157C1 from Streptomyces coelicolor A3(2) which has the motif E(297)QSLW(301) rather than the invariant EXXR motif in the P450 K-helix. Site-directed mutagenesis of native E(297)QSLW(301) in CYP157C1 to E(297)ESLR(301) or E(297)QSRW(301) both containing standard EXXR motifs produced cytochrome P420 proteins thought to be inactive forms of P450 even though wild type CYP157C1 has the spectral properties of a normal P450. These results indicate that the EXXR motif is not required in all CYP tertiary architectures and only a single cysteine residue, which coordinates as the fifth thiolate ligand to the P450 haem iron, is invariant in all CYPs structures. 相似文献
9.
Yelena V. Grinkova 《Biochemical and biophysical research communications》2010,398(2):194-539
Traditional reconstitution of membrane cytochromes P450 monooxygenase system requires efficient solubilization of both P450 heme enzymes and redox partner NADPH dependent reductase, CPR, either in mixed micellar solution or by incorporation in liposomes. Here we describe a simple alternative approach to assembly of soluble complexes of monomeric human hepatic cytochrome P450 CYP3A4 with CPR by co-incorporation into nanoscale POPC bilayer Nanodiscs. Stable and fully functional complexes with different CPR:CYP3A4 stoichiometric ratios are formed within several minutes after addition of the full-length CPR to the solution of CYP3A4 preassembled into POPC Nanodiscs at 37 °C. We find that the steady state rates of NADPH oxidation and testosterone hydroxylation strongly depend on CPR:CYP3A4 ratio and reach maximum at tenfold molar access of CPR. The binding of CPR to CYP3A4 in Nanodiscs is tight, such that complexes with different stoichiometry can be separated by size-exclusion chromatography. Reconstitution systems based on the co-incorporation of CPR into preformed Nanodiscs with different human cytochromes P450 are suitable for high-throughput screening of substrates and inhibitors and for drug-drug interaction studies. 相似文献
10.
Jeffrey G. Scott Padma Sridhar Nannan Liu 《Archives of insect biochemistry and physiology》1996,31(3):313-323
Cytochrome P450tpr is a xenobiotic metabolizing P450 that is found in house flies (Musca domestica). To better understand the regulation of cytochrome P450tpr, the effects of 21 potential monooxygenase inducers were examined for their ability to induce total cytochromes P450 and cytochrome P450tpr levels in adult flies. Six compounds caused induction of total cytochromes P450 per mg protein in adult susceptible (CS) house flies: ethanol (1.6-fold), phenobarbital in food (1.5-fold) or water (1.5-fold), naphthalene (1.3-fold), DDT (1.3-fold), xanthotoxin (1.4-fold), and α-pinene (1.2-fold). Six compounds were found to be inducers of cytochrome P450tpr: piperonyl butoxide in food (1.9-fold), phenobarbital in food (1.4-fold) and water (3.4-fold), clofibrate (1.3-fold), xanthotoxin (1.3-fold), methohexital (1.3-fold), and isosafrole (1.3-fold). Comparison of our results with house fly P450 6A1 indicates that there are specific inducers for each of these individual P450s as well as compounds that induce both P450s. Total P450s were inducible by PB in CS house fly larvae, but not in LPR larvae. Immunoblotting revealed no detectable P450tpr in control or PB-treated larvae in either strain. Thus, although total P450s are inducible in the susceptible strain larvae, P450tpr does not appear to be normally present or inducible with PB in larvae of either strain. Northern blots of phenobarbital (in water) treated CS flies indicated that there was a 4.2-fold increase in the P450tpr (i.e., CYP6D1) mRNA levels over the untreated flies. In the multiresistant LPR strain there was no apparent induction of CYP6D1 mRNA by phenobarbital. Following phenobarbital induction, the level of CYP6D1 mRNA in the CS strain was about half of the level in the LPR strain. © 1996 Wiley-Liss, Inc. 相似文献
11.
昆虫细胞色素P450研究的一些新进展 总被引:5,自引:0,他引:5
报道了有关细胞色素P45 0研究的一些新发现。果蝇和冈比亚按蚊基因组测序的完成 ,使人类对昆虫P45 0的多样性有一完整的概念 ,已查明果蝇和冈比亚按蚊基因组中分别含有 90种和 1 1 1种P45 0基因。P45 0介导的果蝇对DDT的抗性被证明是Cyp6g1基因超量表达的结果。昆虫可以窃听植物分子信号 (水杨酸、茉莉酮酸 ) ,通过P45 0的诱导机制增强自身对植物防御物质的反防御能力。从分子水平上鉴定了 2个参与蜕皮素合成的线粒体P45 0基因。细胞色素P45 0在昆虫信息素降解中的作用得到鉴定。 相似文献
12.
The involvement of cytochrome P450 monooxygenases in methanol elimination in Drosophila melanogaster larvae 总被引:1,自引:0,他引:1
Wang SP He GL Chen RR Li F Li GQ 《Archives of insect biochemistry and physiology》2012,79(4-5):264-275
Methanol is one of the most common short-chain alcohols in fermenting fruits, the natural food of the fruit fly, Drosophila melanogaster. The larvae cope continuously with methanol at various concentrations in order to survive and develop. In the present article, we found toxicities of dietary methanol and formaldehyde were enhanced by piperonyl butoxide, but not by 3-amino-1, 2, 4-triazole, 4-methylpyrazole, diethylmeleate, and triphenyl phosphate, when assessing by the combination index method. These results reveal that cytochrome P450 monooxygenases (CYPs), rather than catalases, alcohol dehydrogenases, glutathione S-transferases, and esterases, participate in methanol metabolism. Moreover, methanol exposure dramatically increased CYP activity. The ratios of the CYP activities in treated larvae to those in control reached, respectively, up to 3.0-, 3.9-, and 2.7-fold, at methanol concentrations of 22.6, 27.9, and 34.5 mg/g diet. In addition, methanol exposure greatly up-regulated the mRNA expression level of five Cyp genes, which were Cyp304a1, Cyp9f2, Cyp28a5, Cyp4d2, and Cyp4e2. Their resulting proteins were suggested as the candidate enzymes for methanol metabolism in D. melanogaster larvae. 相似文献
13.
细胞色素P450(P450s, CYPs)超基因家族是由数量众多、 功能复杂的一类血红蛋白酶基因所组成, 对许多结构多样的外源与内源化合物起着氧化代谢的作用。本文系统地综述了家蚕Bombyx mori基因组中P450s的数量与种类, 其数量比食腐昆虫和杂食性的植食昆虫少, 但比意大利蜜蜂Apis mellifera多, 在基因组中大多呈串联重复排列, 基因结构与系统进化树之间有着紧密的联系。家蚕与黑腹果蝇Drosophila melanogaster P450s的比较基因组学分析发现, 存在10对直向同源基因, 但CYP3与CYP4集团(Clan)的P450s表现出种属特异性扩增。家蚕P450s与其蜕皮激素和保幼激素合成代谢有关, 并涉及到对氟化物与杀虫剂抗性。家蚕作为鳞翅目昆虫的代表, 通过对其P450s的研究, 可望为其他昆虫, 特别是鳞翅目昆虫的生长发育研究和抗性治理提供理论依据和研究模式。 相似文献
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The involvement of cytochrome b5 in different cytochrome P450 monooxygenase and palmitoyl CoA desaturase activities in microsomes from insecticide-resistant (LPR) house flies was determined using a specific polyclonal antiserum developed against house fly cytochrome b5. Anti-b5 antiserum inhibited the reduction of cytochrome b5 by NADH-cytochrome b5 reductase. The antiserum also inhibited palmitoyl CoA desaturase, methoxycoumarin-O-demethylase (MCOD), ethoxycoumarin-O-deethylase (ECOD), and benzo[a]pyrene hydroxylase (aromatic hydrocarbon hydroxylase, AHH) activities. However, methoxyresorufin-O-demethylase (MROD) and ethoxyresorufin-O-deethy-lase (EROD) activities were not affected by this antiserum. These results demonstrate that cytochrome b5 is involved in fatty acyl CoA desaturase activities and in certain cytochrome P450 monooxygenase activities (i.e., MCOD, ECOD, and AHH) in LPR house fly microsomes. Other cytochrome P450 monooxygenase activities (i.e., MROD and EROD) may not require cytochrome b5. The results suggest that cytochrome b5 involvement with cytochrome P450 monooxygenase activities is dependent upon the cytochrome P450 isoform involved. © 1994 Wiley-Liss, Inc. 相似文献
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Mohammad Asaduzzaman Miah Mohammed Esmail Abdalla Elzaki Asmaul Husna Zhaojun Han 《Archives of insect biochemistry and physiology》2019,100(2)
Deltamethrin resistance in Laodelphax striatellus had been associated with its oxidative detoxification by overexpression of four cytochrome P450 monooxygenases like CYP353D1v2, CYP6FU1, CYP6AY3v2, and CYP439A1v3. The first three P450s have been validated for insecticide‐metabolizing capability and only CYP6FU1 was found to degrade deltamethrin. In this study, an investigation was conducted to confirm the capability of CYP439A1v3 to degrade deltamethrin. The CYP439A1v3 was first expressed in Sf9 cell line and its recombinant enzyme was tested for metabolic activity against different insecticides using substrate depletion assay combined with metabolite identification. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and carbon monoxide (CO)‐difference spectra analysis showed that the intact cytochrome P450 protein was successfully expressed. Tests with probe substrates proved its enzyme activity, as p‐nitroanisole, ethoxycoumarin, and ethoxyresorufin were preferentially metabolized (specific activity 7.767 ± 1.22, 1.325 ± 0.37, and 0.355 ± 0.37 nmol/min per mg of protein, respectively) while only luciferin‐HEGE was not. In vitro incubation of the recombinant CYP439A1v3 protein with deltamethrin revealed hydroxylation by producing hydroxydeltamethrin. On the contrary, no metabolite/metabolism was seen with nonpyrethroid insecticide, including imidacloprid, buprofezin, chlorpyrifos, and fipronil. To the best of our knowledge, this is the first study to link a CYP450 from family 439 to confer pyrethroid resistance to L. striatellus. This finding should help in the design of appropriate insecticide resistance management for control of this strain of L. striatellus. 相似文献
16.
《Current biology : CB》2022,32(3):671-676.e5
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17.
David R. Nelson Jared V. Goldstone John J. Stegeman 《Philosophical transactions of the Royal Society of London. Series B, Biological sciences》2013,368(1612)
The neighbourhoods of cytochrome P450 (CYP) genes in deuterostome genomes, as well as those of the cnidarians Nematostella vectensis and Acropora digitifera and the placozoan Trichoplax adhaerens were examined to find clues concerning the evolution of CYP genes in animals. CYP genes created by the 2R whole genome duplications in chordates have been identified. Both microsynteny and macrosynteny were used to identify genes that coexisted near CYP genes in the animal ancestor. We show that all 11 CYP clans began in a common gene environment. The evidence implies the existence of a single locus, which we term the ‘cytochrome P450 genesis locus’, where one progenitor CYP gene duplicated to create a tandem set of genes that were precursors of the 11 animal CYP clans: CYP Clans 2, 3, 4, 7, 19, 20, 26, 46, 51, 74 and mitochondrial. These early CYP genes existed side by side before the origin of cnidarians, possibly with a few additional genes interspersed. The Hox gene cluster, WNT genes, an NK gene cluster and at least one ARF gene were close neighbours to this original CYP locus. According to this evolutionary scenario, the CYP74 clan originated from animals and not from land plants nor from a common ancestor of plants and animals. The CYP7 and CYP19 families that are chordate-specific belong to CYP clans that seem to have originated in the CYP genesis locus as well, even though this requires many gene losses to explain their current distribution. The approach to uncovering the CYP genesis locus overcomes confounding effects because of gene conversion, sequence divergence, gene birth and death, and opens the way to understanding the biodiversity of CYP genes, families and subfamilies, which in animals has been obscured by more than 600 Myr of evolution. 相似文献
18.
细胞色素P450酶系广泛分布于各种生物中,它们通常由一组基因超家族编码并含有血红素,能够催化一系列化学反应,具有多种生物学功能。特别是原核生物P450酶在催化内源性和外源性化合物的反应中具有重要的工业生产应用价值,成为近年来P450酶系研究的热点。本文对近年来原核生物P450酶系的重组表达和生物催化领域的研究进展进行综述。 相似文献
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Insight into the redox partner interaction mechanism in cytochrome P450BM‐3 using molecular dynamics simulations
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Flavocytochrome P450BM‐3 is a soluble bacterial reductase composed of two flavin (FAD/FMN) and one HEME domains. In this article, we have performed molecular dynamics simulations on both the isolated FMN and HEME domains and their crystallographic complex, with the aim to study their binding modes and to garner insight into the interdomain electron transfer (ET) mechanism. The results evidenced an interdomain conformational rearrangement that reduces the average distance between the FMN and HEME cofactors from 1.81 nm, in the crystal structure, to an average value of 1.41 ± 0.09 nm along the simulation. This modification is in agreement with previously proposed hypotheses suggesting that the crystallographic FMN/HEME complex is not in the optimal arrangement for favorable ET rate under physiological conditions. The calculation of the transfer rate along the simulation, using the Pathways Path method, demonstrated the occurrence of seven ET pathways between the two redox centers, with three of them providing ET rates (KET) comparable with the experimental one. The sampled ET pathways comprise the amino acids N319, L322, F390, K391, P392, F393, A399, C400, and Q403 of the HEME domain and M490 of the FMN domain. The values of KET closer to the experiment were found along the pathways FMN(C7) → F390 → K391 → P392 → HEME(Fe) and FMN(C8) → M490 → F393 → HEME(Fe). Finally, the analysis of the collective modes of the protein complex evidences a clear correlation of the first two essential modes with the activation of the most effective ET pathways along the trajectory. © 2013 Wiley Periodicals, Inc. Biopolymers 101: 197–209, 2014. 相似文献