Ahpf protein. AlphaFold 2 Protein Structure Predictions.
Ahpf protein For improvements in protein expression and hydro-peroxide sensitivity testing, ahpC, ahpF, and the The present invention relates to an AhpF protein which has thioredoxin reductase, peroxidase, and chaperone activities and is derived from Pseudomonas aeruginosa, and a use therefor. To complete the catalytic cycle, the oxidized AhpCs are reduced by alkyl hydroperoxide reductase AhpD, AhpF, or thioredoxin C (TrxC) (Bryk et al. Functionally, AhpC and AhpE considered as peroxidases directly detoxify peroxides, while AhpD and AhpF as oxidoreductases restore oxidized peroxidases to their reduced form. However, how Ahp deficiency-induced ROS modulate the redox imbalance and metabolic alterations atill warrant investigation and needs to be further A chimeric protein with the first 207 amino acids of S. The protein consists of an N-terminal domain (NTD), which catalyzes the reduction of redoxactive disulfides in AhpC [10,11] (Fig. 4. Links. Here we demonstrate that streptomycin, a bactericidal aminoglycoside that increases ribo … Purified Recombinant Bacillus subtilis AHPF protein, His-tagged from Creative Biomart. , 2004; Wong et al. Catalytic activity. AhpF is a 57 kDa dimeric flavoprotein that utilizes reducing equivalents from NAD-(P)H to reduce its substrate protein, AhpC (a member of the peroxiredoxin family). microorganisms (Link et al. , 2004; Koshkin et al. radiodurans [4]. 2-D gels. coli Trx were expressed and purified as previously described (6, 17, 18). 11. Many eubacterial genomes including those of Salmonella typhimurium, Streptococcus mutans, and Thermus aquaticus encode a dedicated flavoprotein reductase (AhpF, Nox1, or PrxR) just downstream of the structural gene for their peroxiredoxin (Prx, AhpC) homologue to reduce the latter protein during turnover. No ahpF gene appears to be present, although a in S. tuberculosis which encodes AhpE, AhpD is known to substitute for AhpF [17]. In this study we report the identification and role of the alkyl hydroperoxide reductase (ahp) gene in Bacteroides fragilis. The process was performed using the The genes encoding several of these proteins, including the ferric uptake regulatory protein (Fur), the alkyl hydroperoxide reductase (AhpR) subunits F (AhpF) and C (AhpC), and the DNA protection during starvation protein (Dps), are under the control of the OxyR transcriptional dual regulator. The AhpR system, composed of AhpC and a flavoprotein reductase, AhpF, catalyzes the pyridine nucleotide-dependent reduction of organic hydroperoxides and Feb 29, 2008 · The gene products of ahpC and ahpF share 89% and 75% sequence identity with AhpC (BAA25695) and AhpF (BAA25696) of Streptococcus mutans, respectively . , 1997; Wood et al. 8 mM Dec 1, 2022 · total protein sequences for AhpF (Table 2). typhimurium was included in place of TrxR-Trx1 with H. 55 µg of protein from pulse-labeled samples for analytical gels and 300 The invention relates to an AhpF protein derived from Pseudomonas aeruginosa (Pseudomonas aeruginosa) and application thereof, wherein the AhpF protein has thioredoxin reductase, peroxidase and molecular chaperone activities. 26) reduction (Hong et al. By using a novel activity of the AhpF of Pseudomonas aeruginosa according to the present invention, it is possible to produce a plant having strong resistance to various environmental stresses such as oxidative stress or heat stress, thereby making it Legend. Although AhpD has long been demonstrated in Mycobacterium tuberculosis, its physi … An AhpF protein has thioredoxin reductase, peroxidase, and chaperone activities and is derived from Pseudomonas aeruginosa, and a use therefor. May 5, 2000 · AhpF, the flavin-containing component of the Salmonella typhimurium alkyl hydroperoxide reductase system, catalyzes the NADH-dependent reduction of an active-site disulfide bond in the other component, AhpC, which in turn reduces hydroperoxide substrates. It was found the concentration of endogenous H2O2 was submicromolar in a mutant strain E. 0 Å and Circular permutation was found to be a feature of the AhpF protein family. Dec 25, 2001 · Indeed, the AhpF-derived N-terminal redox center was reduced by the thioredoxin reductase part of the chimeric molecule during anaerobic dithionite or NADPH titrations, and the chimeric protein exhibited a level of catalytic efficiency in peroxidase assays with NADPH and AhpC nearly identical to that of AhpF itself . Feb 25, 2021 · Abstract Alkyl hydroperoxide reductase (AhP), catalase G (KatG), and catalase E (KatE) are the main enzymes to scavenge the excessive hydrogen peroxide in E. 2019). fragilis AhpCF shares approximately 60% identity t … cd03026 (PSSM ID: 239324): Conserved Protein Domain Family AhpF_NTD_C, TRX-GRX-like family, Alkyl hydroperoxide reductase F subunit (AhpF) N-terminal domain (NTD) subfamily, C-terminal TRX-fold subdomain; AhpF is a homodimeric flavoenzyme which catalyzes the NADH-dependent reduction of the peroxiredoxin AhpC, which then reduces hydrogen peroxide and organic hydroperoxides Oct 22, 2019 · The presence of AhpC in the protein extract might sequester the AhpF protein, precluding its hypothesized nitrofuran reductase activity. Settings. , 2002; Jaeger et al. doi: 10. The transcriptional start sites of katB-ankB, ahpB, and ahpC-ahpF were mapped, putative OxyR-binding sites were identified upstream of the -35 promoter elements, and direct binding of purified OxyR protein to these target promoters was demonstrated. E. An AhpF protein has thioredoxin reductase, peroxidase, and chaperone activities and is derived from Pseudomonas aeruginosa , and a use therefor. AhpF is one of the ten. AhpF, a flavoprotein with two redox-active disulfide centers per subunit (Cys129Cys132 and Cys345Cys348), Sep 4, 2019 · Introduction. tuberculosis genome has been sequenced in its en-tirety (25). 9 and then induced for protein expression by addition of IPTG to a final concentration of 0. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed. A candidate for such an AhpF equivalent is thioredoxin reductase. Assays of TrxR-Trx1 excluding the AhpC protein (dashed-dotted line) and assays of TrxR alone (small dashes) were also conducted. 5 μM) from S. Source: COG: Taxonomy: Bacteria: Protein: Representatives: Specific Jan 7, 2020 · Alkyl hydroperoxide reductase subunit F (AhpF) is a well-known flavoprotein that transfers electrons from pyridine nucleotides to the peroxidase protein AhpC via redox-active disulfide centers to detoxify hydrogen peroxide. Reduced AhpC is a direct reductant of organic hydroperoxides and hydrogen peroxide (3, 4). Dec 14, 2012 · Protein mistranslation causes growth arrest in bacteria, mitochondrial dysfunction in yeast, and neurodegeneration in mammals. coli reduces the oxidative damage of mistranslated proteins, alleviates the cellular toxicity of protein misfolding, and improves bacterial fitness in the presence of aminoglycosides. *) are essential for the oxygen tolerance of aerobic organisms by converting otherwise toxic hydroperoxides of lipids or nucleic acids to the corresponding alcohols. 197/0. The present invention relates to an AhpF protein which has thioredoxin reductase, peroxidase, and chaperone activities and is derived from Pseudomonas aeruginosa , and a use therefor. The katB , ahpB , and ahpC-ahpF loci are shown with their coordinates in the PAO1 genome (Pathogenesis Corp. Jul 1, 2000 · AhpF of Salmonella typhimurium, the flavoprotein reductase required for catalytic turnover of AhpC with hydroperoxide substrates in the alkyl hydroperoxide reductase system, is a 57 kDa protein with homology to thioredoxin reductase (TrR) from Escherichia coli. Apr 5, 1999 · In this study we report the identification and role of the alkyl hydroperoxide reductase (ahp) gene in Bacteroides fragilis. most abundant proteins in E. typhimurium AhpF attached to the N-terminus of Escherichia coli thioredoxin reductase exhibits very high NADPH:peroxiredoxin oxidoreductase and thioredoxin reductase activities. For the SwissDock server (35, 71), a blind docking simulation was implemented using ready-to-dock FZ ligand data file from ZINC database (accession number ZINC113418) and the AhpF structural data file from Protein Data Bank (PDB identifier [ID] 4O5Q) . Induced cultures were allowed to continue growing for 3 h prior to harvest. faecalis (V583) AhpF (EfAhpF) appears to be a representative of a minor subclass of this family, the typically N-terminal two-fold thioredoxin-like domain (NTD_N/C) is located at the C-terminus, whereas the pyridine nucleotide-disulfide oxidoreductase domain is ahpf protein pseudomonas aeruginosa activity ahpf protein Prior art date 2013-09-17 Application number PCT/KR2013/010564 Other languages English (en) French (fr) Korean (ko) Inventor 정병엽 싱수디르 이승식 배형우 이성범 Original Assignee 한국원자력연구원 Priority date (The priority date is an assumption and is not a Apr 21, 2017 · AhpF is a flexible multidomain protein that enables a series of electron transfers among the redox centers by accepting reducing equivalents from NADH. coli MG1655/ΔAhpΔKatEΔKatG, which was enough to cause damage to DNA and proteins as well as concomitant cell growth and metabolism. S. AhpF (0. AhpF is one of the ten most abundant proteins in E. The remaining catalytically inactive Cys476 and Cys489 of AhpF and Cys105 and Cys303 of TrR are not Nov 1, 2023 · Additionally, overexpression of AhpF has been found to suppress protein aggregation upon antibiotic stimulation, suggesting that cellular defense against H 2 O 2 can reduce the cell damage [13]. 88% of the total protein sequences for AhpF (Table 2). Fractions containing protein are pooled, dialyzed in storage buffer (200 mM NaCl, 20 mM Tris HCl pH 7. It remains poorly understood how mistranslated proteins cause such cellular defects. radiodurans possesses an AhpD-like protein DR1765 (COG2128) which is also observed in all other Deinococcus species [4]. The two components of ahp, ahpC, and ahpF, are organized in an operon, and the deduced amino acid sequences revealed that B. AhpD recycles AhpC by linking dihydrolipoamide dehydrogenase (Lpd) and Protein was purified using a HiTrap IMAC FF 1 mL column (GE Healthcare), eluting with 1 M Imidazole. Two-dimensional gel electrophore … The present invention relates to: an AhpF protein which has a thioredoxin reductase, peroxidise, and chaperone activity and is derived from Pseudomonas aeruginosa , and to a use therefor. AhpF contains the reductant (NAD(P)H) binding domain (pfam00070) and presumably acts to resolve the disulfide which forms after oxidation of the active site cysteine in AphC. Rhea 62628. Because the mutant protein still functions as an electron acceptor from AhpF, the electrons necessary to restore growth to the trxB,gor strain must be derived from NADH. Other Protein Purifications. Not only does the C129,C132 disulfide center of this domain carry out a Trx-like function in mediating electron transfer from the TrxR-like part of AhpF to its protein disulfide substrate, Jul 11, 2017 · Instead of AhpF, AhpC is reduced by (Fig. Apr 4, 2017 · AhpF is a homodimeric protein encompassed by an N-terminal domain (NTD) and a C-terminal domain (CTD), which includes an FAD- and NADH-binding site as well as the Oct 28, 1997 · The catalytic properties of cysteine residues Cys46 and Cys165, which form intersubunit disulfide bonds in the peroxidatic AhpC protein of the alkyl hydroperoxide reductase (AhpR) system from Salmonella typhimurium, have been investigated. Reactive oxygen species (ROS) are produced by aerobic respiration and immune defense of organisms (Lü et al. action of a 57 kDa FAD-containing protein, AhpF, and a 21 kDa protein, AhpC (a member of the peroxiredoxin family) (1, 2). Characterization of the purified AhpF and AhpC proteins has clarified the role of each in catalysis of peroxide reduction. In this proposed scheme, AhpF transfers electrons from reduced pyridine nucleotides to AhpC Via the flavin and direct dithiol-disulfide interchange, followed by transfer of electrons from the dithiol center of AhpC to the hydroperoxide substrate. 2020), while AhpD assumes responsibility for reducing oxidized AhpC (EC: 1. May 5, 2008 · Sulfiredoxin (Srx) is one of a family of low molecular weight sulfur containing proteins linked with maintenance of cellular redox balance. Data collection and structure determination of Ec AhpF. Structure, mechanism and ensemble formation of the alkylhydroperoxide reductase subunits AhpC and AhpF from Escherichia coli Acta Crystallogr D Biol Crystallogr . coli AhpF protein and a three-wavelength multiwavelength anomalous diffraction (MAD) data set for the dysprosium derivative were collected at 140 K on beamline 13B1 at the National Synchrotron Radiation Research Center (NSRRC), Hsinchu, Taiwan using an ADSC Quantum 315 CCD detector. These proteins form a cysteine-based bicomponent H 2 O 2 -decomposing complex, as shown by in vitro studies with recombinant proteins ( 32 ). chimeric protein with the first 207 amino acids of S. The amino acid sequence of the C-terminus of … Pseudomonas aeruginosa PAO1, PA0140 (ahpF) Cytoplasmic Cytoplasmic Membrane Periplasmic Outer Membrane Extracellular AlphaFold 2 Protein Structure Predictions. coli, and is also present in other. C. 4 mM. 5, 1 mM EDTA, 0. Spectral properties of the the wild type and double cysteine mutant AhpF proteins. An AhpF protein has thioredoxin reductase, peroxidase, and chaperone activities and is derived from Pseudomonas aeruginosa, and a use therefor. coelicolor and Mycobacterium spp. tuberculosis appears to lack AhpF (Wilson and Collins 1996; and this work), so that for anti-RNI defense, another protein may shuttle electrons from NAD(P)H to AhpC in place of AhpF. 2. Oct 28, 1997 · AhpF, the alkyl hydroperoxide reductase component which transfers electrons from pyridine nucleotides to the peroxidase protein, AhpC, possesses two redox-active disulfide centers in addition to one FAD per subunit; the primary goal of these studies has been to test for the requirement of one or both of these disulfide centers in catalysis. Homology of AhpF to the smaller (35 kDa) TrR protein occurs in the C-terminal part of AhpF; a stretch of about 200 amino acids at the N-terminus of AhpF 2-Cys peroxiredoxins (Prxs) are a large family of peroxidases, responsible for antioxidant function and regulation in cell signaling, apoptosis and differentiation. Due to the influence of evolutionary levels, gene replication, horizontal Sep 1, 2000 · Genetic maps and transcripts of OxyR-regulated genes. , 2001; Jiang and. 1). Alkyl hydroperoxide reductase subunit F (AhpF) is a well-known flavoprotein that transfers electrons from pyridine nucleotides to the peroxidase protein AhpC via redox-active disulfide centers to detoxify hydrogen peroxide. typhimurium AhpF, E. Considerable evidence now strongly supports this general scheme. Protein amount was determined by Bradford assay using BSA as Jun 17, 2015 · AhpF is a flexible multi-domain protein that enables a series of electron transfers among the redox centers by accepting reducing equivalents from NADH. by the AhpF system in bacteria or by the thioredoxin/ thioredoxin reductase system in animals and fungi. Surprisingly, this chimeric The present invention relates to: an AhpF protein which has a thioredoxin reductase, peroxidise, and chaperone activity and is derived from Pseudomonas aeruginosa, and to a use therefor. , 2017). For improvements in protein expression and hydro-peroxide sensitivity testing, ahpC, ahpF, and the Aug 20, 2020 · In addition, there is no AhpF homologue in D. By using a novel activity of the AhpF of Pseudomonas aeruginosa according to the present invention, it is possible to produce a plant having strong resistance to various environmental stresses such as oxidative stress or heat stress, thereby making it Mar 16, 2001 · Alkylhydroperoxide reductases (AhpR, EC 1. coelicolor and Mycobacterium species, the ahpF gene is absent, whereas downstream of the ahpC gene is located the ahpD gene, which encodes a thioredoxin-like protein (19 kDa) involved as electron donor in the disulfide reduction of AhpC. Oct 8, 2024 · Protein biosynthesis rates were determined by measuring the incorporation of radioactive methionine in auranofin-treated and untreated samples after protein precipitation with 20% (wt/vol) trichloro acetic acid using a scintillation counter (Tri-Carb 2800TR, PerkinElmer). Creative BioMart to Present at BPS 2025 Annual Meeting | February 15-19, 2025 Oct 28, 1997 · AhpF, the alkyl hydroperoxide reductase component which transfers electrons from pyridine nucleotides to the peroxidase protein, AhpC, possesses two redox-active disulfide centers in addition to one FAD per subunit; the primary goal of these studies has been to test for the requirement of one or bot … Dec 25, 2001 · Indeed, the AhpF-derived N-terminal redox center was reduced by the thioredoxin reductase part of the chimeric molecule during anaerobic dithionite or NADPH titrations, and the chimeric protein exhibited a level of catalytic efficiency in peroxidase assays with NADPH and AhpC nearly identical to that of AhpF itself . By similarity. Creative BioMart to Present at IMMUNOLOGY2024™|May 3-7, 2024|Booth #512 Learn More DVU0283 AhpF family protein/thioredoxin reductase [] Gene ID: 2796693, discontinued on 6-Mar-2020. In the presence of the 22-kDa colorless component (AhpC) of the Salmonella alkyl-hydroperoxide reductase, both proteins catalyze the 4-electron reduction of oxygen to water. It can use either NADH or NADPH as electron donor for direct reduction of redox dyes or of alkyl hydroperoxides when combined with the AhpC protein. ) Abandoned Application number US15/022,296 Other languages English (en Apr 3, 2001 · AhpF, a homodimer of 57 kDa subunits, is a flavoenzyme which catalyzes the NADH-dependent reduction of redox-active disulfide bonds in the peroxidase AhpC, a member of the recently identified peroxiredoxin class of antioxidant enzymes. 55 µg of protein from pulse-labeled samples for analytical gels and 300 Salmonella disrupted in ahpC became hypersusceptible to RNI. The structure of AhpF from Salmonella typhimurium at 2. It also contains a catalytic core similar to TRX reductase containing FAD and NADH binding domains with an active site disulfide. Protein-protein interaction databases. 1B) as well as a protein sequence analysis of mycobacterial AhpCs revealed differences in the amino acid composition of AhpCs from other sources Mar 7, 2001 · AhpF, a homodimer of 57 kDa subunits, is a flavoenzyme which catalyzes the NADH-dependent reduction of redox-active disulfide bonds in the peroxidase AhpC, a member of the recently identified peroxiredoxin class of antioxidant enzymes. Protein AhpF (wt)a C132S/C348S C132S/C345S C129S/C348S C129S/C345S Molar extinction coefficient b 13,000 ± 660 12,020 ± 200 11,360 ± 120 11,050 ± 120 11,200 ± 250 λmax of two peaks 381, 449 374, 447 382, 449 374, 447 382, 449 AhpF also catalyzes NADH-dependent hydrogen peroxide formation under aerobic conditions, albeit at a somewhat slower rate than the Amphibacillus protein. typhimurium fully complemented the defect. The M. , 12-15-99 release), the sites of An AhpF protein has thioredoxin reductase, peroxidase, and chaperone activities and is derived from Pseudomonas aeruginosa, and a use therefor. However, few studies explored how submicromolar intracellular hydrogen peroxide alters protein function and regulates the signaling pathways at the proteome level. the wild type and was not detectable in an ahpC mutant, presumably Jun 1, 2007 · AhpF, a separate disulfide reductase protein, regenerates AhpC every catalytic cycle via electrons from NADH which are transferred to AhpC through a tightly bound flavin and two disulfide centers The activity in these assays depends on the protein concentration, as shown in Fig. This proteins contains two paired conserved cysteine motifs, CxxCP and CxHCDGP. Feb 12, 2002 · Many eubacterial genomes including those of Salmonella typhimurium, Streptococcus mutans, and Thermus aquaticus encode a dedicated flavoprotein reductase (AhpF, Nox1, or PrxR) just downstream of the structural gene for their peroxiredoxin (Prx, AhpC) homologue to reduce the latter protein during tur … The AhpD protein is about 19 kDa, much smaller than AhpF (52 kDa), and does not contain FAD or NAD(P)H binding domains conserved in AhpF proteins. AhpF, a homo-dimer of 57 kDa subunits, is an FAD-dependent NADH: protein-disulfide reductase which reduces redox-active di-sulfides in AhpC during catalytic turnover (8, 9). Dec 14, 2012 · In summary, we propose that overexpression of AhpF in E. fragilis AhpCF shares approximately 60% identity to orthologues in other gram-positive and gram-negative bacteria. A single-wavelength data set for the native E. Jan 9, 1996 · The two components, AhpF and AhpC, of the Salmonella typhimurium alkyl hydroperoxide reductase enzyme system have been overexpressed and purified from Escherichia coli for investigations of their catalytic properties. Jan 1, 2016 · The determined EcAhpC-AhpF solution ensemble with a stoichiometry of AhpC 10:AhpF 2 reveals that the extended conformation of the dimeric EcAhpF brings the Cys–Cys centers of the NTD in neighborhood to the redox-active disulphide centers of EcAhpC and provides an efficient electron transfer between EcAhpF and the catalytic EcAhpC protein. Unlike protection against cumene hydroperoxide, protection afforded by ahpC against RNI was independent of the reducing flavoprotein, AhpF. ahpC from either Mycobacterium tuberculosis or S. 1. 2009; Sosa et al. Alkyl hydroperoxide reductase subunit F (AhpF) is a well-known flavoprotein that transfers electrons from pyridine nucleotides to the peroxidase protein AhpC via redox-active disulfide centers to detoxify hydrogen peroxide. bearing the entire coding region for AhpF (pBlF2; 4; Figure Scheme 1 FIGURE 1: Location of the tryptic cleavage site and redox-active half-cystine residues within the AhpF protein sequence in com-parison with those of thioredoxin reductase (TrR). ) Abandoned Application number US15/022,296 Other languages English (en In silico docking of furazolidone to the AhpF protein. May 1, 1998 · Defense against RNI does not involve AhpF, and M. Fractions containing pure Cp20 were pooled, dialyzed against Buffer A, and concentrated prior to storage at -20 °C. A flexible linker connecting the N-terminal Jun 23, 2000 · The activity in these assays depends on the protein concentration, as shown in Fig. coli cells were lysed on ice in buffer A (50 mM Tris–HCl pH 8, 500 mM NaCl, 2 mM PMSF, 1 mM Pefabloc SC, 0. Protein AhpF (wt)a C132S/C348S C132S/C345S C129S/C348S C129S/C345S Molar extinction coefficient b 13,000 ± 660 12,020 ± 200 11,360 ± 120 11,050 ± 120 11,200 ± 250 λmax of two peaks 381, 449 374, 447 382, 449 374, 447 382, 449 Table S2. Nox-1 is homologous with the flavoprotein component, AhpF, of Salmonella typhimurium alkyl hydroperoxide reductase. In Mycobacteria, AhpC is reduced by AhpD instead of AhpF. Recombinant proteins were isolated in high yield (25-33 mg per liter of bacterial … ahpf protein, recombinant ahpf protein. 2010). coli AhpF E. By using a novel activity of the AhpF of Pseudomonas aeruginosa according to the present invention, it is possible to produce a plant having strong resistance to various environmental stresses such as oxidative stress or heat stress, thereby making it Aug 1, 2000 · Like TrR, AhpF employs tightly bound FAD and redox-active disulfide center(s) in catalyzing electron transfer from reduced pyridine nucleotides to the disulfide bond of its protein substrate. 3. 2014 Nov;70(Pt 11):2848-62. No AhpF-homologuous enzymes have been described for eukaryotes. Catalytic turnover by NADH:peroxiredoxin oxidoreductases may involve major domain Jan 1, 2016 · The determined EcAhpC-AhpF solution ensemble with a stoichiometry of AhpC 10:AhpF 2 reveals that the extended conformation of the dimeric EcAhpF brings the Cys–Cys centers of the NTD in neighborhood to the redox-active disulphide centers of EcAhpC and provides an efficient electron transfer between EcAhpF and the catalytic EcAhpC protein. 217 for data between 10. Further refinement was carried out using CNS 21 until the refinement converged at a R-factor/R-free of 0. Jun 6, 2000 · AhpF, the flavin-containing component of the Salmonella typhimurium alkyl hydroperoxide reductase system, catalyzes the NADH-dependent reduction of an active-site disulfide bond in the other component, AhpC, which in turn reduces hydroperoxide substrates. Catalytic turnover by NADH:peroxiredoxin oxidoreductases may involve major domain The genes encoding several of these proteins, including the ferric uptake regulatory protein (Fur), the alkyl hydroperoxide reductase (AhpR) subunits F (AhpF) and C (AhpC), and the DNA protection during starvation protein (Dps), are under the control of the OxyR transcriptional dual regulator. 4259893 72 interactors; ahpf protein pseudomonas aeruginosa ahpf protein plant Prior art date 2013-09-17 Legal status (The legal status is an assumption and is not a legal conclusion. In order to study the effect of endogenous oxidative stress caused by submicromolar hydrogen peroxide, this study first constructed a mutant strain E. typhimurium AhpF), to the third, Trx-like domain at the N-terminus of AhpF (blue domain in Figure 1). Oct 5, 2001 · These results suggest that the mutant AhpC* protein, although it has acquired a new reductive function, has effectively lost its original ability to reduce peroxides. AhpF is comprised of three domains: an N-terminal redox-active disulfide-containing domain (NTD), an FAD binding. By using a novel activity of the AhpF of Pseudomonas aeruginosa according to the present invention, it is possible to produce a plant having strong resistance to various environmental stresses such as oxidative stress or heat stress, thereby making it of one subunit of F[208-521] and one subunit of AhpFC345,348S; HET 2, heterodimer consisting of one subunit of F[208-521]C345,348S and one subunit of wild type AhpF; DTNB, 5,5′-dithiobis(2-nitrobenzoate); TNB, 2-nitro-5-thiobenzoate; DMSO, dimethyl sulfoxide; FPLC, fast protein liquid chromatography; EDTA, ethylenediaminetetraacetic acid; The present invention relates to an AhpF protein which has thioredoxin reductase, peroxidase, and chaperone activities and is derived from Pseudomonas aeruginosa , and a use t NOVEL ACTIVITY OF AHPF PROTEIN OF PSEUDOMONAS AERUGINOSA, AND USE THEREFOR - KOREA ATOMIC ENERGY RESEARCH INSTITUTE Nov 10, 2014 · Proteomics analysis of the C7 fraction revealed 4 unique peptides covering 12. However, saturation of the AhpD activity only occurred above a 6:1 ratio of the two proteins. Not only does the C129,C132 disulfide center of this domain carry out a Trx-like function in mediating electron transfer from the TrxR-like part of AhpF to its protein disulfide substrate, AhpF contains an NTD forming two contiguous TRX-fold subdomain similar to Pyrococcus furiosus protein disulfide oxidoreductase (PfPDO). coli TrxR, and E. Mar 16, 2001 · The high quality of the 1. In M. 28) in Corynebacterium glutamicum (Hong et al. However Table S2. Oct 21, 2024 · For instance, Pseudomonas aeruginosa employs AhpF as an auxiliary protein to facilitate AhpC (EC: 1. Expression and Purification of Recombinant AhpC and AhpF. One function of Srx is the reduction of cysteine Jan 1, 2001 · Nox-1 from Streptococcus mutans, the bacteria which cause dental caries, was previously identified as an H2O2-forming reduced nicotinamide adenine dinucleotide (NADH) oxidase. Analysis Protein purification of E. Summary A600 value of 0. coli. coli MG1655/ΔAhpΔKatEΔKatG. This protein was induced coordinately with AhpC in. 0 Å resolution, determined using multiwavelength anomalous dispersion, shows that the C-terminal portion Salmonella disrupted in ahpC became hypersusceptible to RNI. 6 ) was performed in the presence of 5 μg/ml of nickel-nitrilotriacetic acid (Ni-NTA) affinity-purified AhpF protein, NADH, and one of the three 5-nitrofurans (FZ, NIT, and NFZ To study the impact of nutritional factors on protein expression of intestinal bacteria, gnotobiotic mice monoassociated with Escherichia coli K-12 were fed three different diets: a diet rich in starch, a diet rich in nondigestible lactose, and a diet rich in casein. coli AhpF. in S. Reductive titrations, catalytic effects of thiol-modify-ing agents, and functional studies of cysteine-to-serine mutants of both proteins strongly support a flavin and redox-active cystine-based mechanism involving AhpF Biochemistry, 2002. Corynebacterium May 21, 2019 · Alkyl hydroperoxidase reductase AhpD, which is functionally equivalent to the bacterial flavin-containing disulfide reductase AhpF, acts as a proton donor for the organic peroxide-scavenging alkyl hydroperoxidase AhpC. Thus, in yeast the alkylhydroperoxidase is coupled to thioredoxin and thioredoxin reductase rather than to an AhpF-like protein (24). A partia … AhpF is a homo-dimeric protein and belongs to FAD-dependent NADH protein-disulfide reductases. Recombinant Bacillus subtilis AHPF protein, His-tagged can be used for research. 6. 0 A resol … Oct 22, 2019 · Using genome sequencing, genetic, biochemical, and bioinformatic approaches, we discovered a novel 5-nitrofuran-activating enzyme, AhpF, in E. 7 for the AhpF assay, and for AhpC was saturable at or above a 3:1 ratio of AhpC:AhpF. Miscellaneous The active site is a redox-active disulfide bond. The excessive amount of ROS can result in cellular damage, which promotes chronic diseases, such as cardiovascular diseases, diabetes, and cancer (Stephens et al. However, study of AhpF has historically been limited to particular eubacteria, and the connection between the functional and structural properties of AhpF remains unknown Sep 20, 2018 · Regeneration of the intermolecular disulphide bond occurs via AhpF, a dedicated AhpC reductase in most bacteria, or thioredoxin, protein oxidation and chaperoning function 12,13,14. BioGRID. However TIGR03143 (PSSM ID: 132187): Conserved Protein Domain Family AhpF_homolog, This family of thioredoxin reductase homologs is found adjacent to alkylhydroperoxide reductase C subunit predominantly in cases where there is only one C subunit in the genome and that genome is lacking the F subunit partner (also a thioredcxin reductase homolog) that is usually found (TIGR03140) May 15, 2007 · AhpF, a separate disulfide reductase protein, regenerates AhpC every catalytic cycle via electrons from NADH which are transferred to AhpC through a tightly bound flavin and two disulfide centers, Cys345-Cys348 and Cys129-Cys132, through putative large domain movements. Aug 22, 2023 · Alkyl hydroperoxide reductase (Ahp), comprised of four different subunits AhpC, AhpD, AhpE, and AhpF, is a thiol-based antioxidative enzyme with the ability to protect bacteria against oxidative stress. The proposed mechanism of action of AhpF is similar to a TRX/TRX reductase system. D. Surprisingly, this chimeric A600 value of 0. In S. 9 Å 2F obs − F calc electron density map allowed semi-automatic tracing and model rebuilding by WARP 20 and was continuous for the entire protein moiety of the E. 2013). coli The discovery of a new nitrofuran-reducing enzyme opens new avenues for overcoming 5-nitrofuran resistance, such as designing nitrofuran analogues with higher affinity for AhpF or screening for ahpf protein pseudomonas aeruginosa ahpf protein plant Prior art date 2013-09-17 Legal status (The legal status is an assumption and is not a legal conclusion. suggests their function as thioredoxin-like proteins involved in AhpR proteins; either protein independently overexpressed in Escherichia coli is partially protective (4). coli, and cently described peroxiredoxin family (6, 7). 1% Triton X-100, 50% Glycerol), and loaded on a SDS-PAGE gel. Alkyl hydroperoxide reductase subunit AhpF [Defense mechanisms] linked to 3D-structure. 1107/S1399004714019233. pasteurianum rubredoxin was purified as reported previously (12). The two components of ahp, ahpC, and ahpF, are organized in an operon, and the deduced amino acid sequences revealed that B. pylori AhpC (medium dashes), and in this case, NADH rather than NADPH was used as the reducing substrate under anaerobic conditions. A flexible linker connecting the N-terminal domain (NTD) and C-terminal domain (CTD) of AhpF suggests that the enzyme adopts a large-scale domain motion that alternates between the closed and Dec 1, 1996 · the 54-kDa protein was identified as the AhpF protein on the. The Escherichia coli alkylhydroperoxide reductase (AhpR) is a prototype of the Prxs-family, and is composed of an NADH-dependent AhpF r … Jan 23, 2007 · The disulfide is subsequently reduced by AhpF. The AhpF component belongs to the family of pyridine nucleotide-disulphide oxidoreductas … The protein required to reduce the AhpC disulfide bond is not the same in all organ-isms. However, the conservation of cysteine residues in the C-X-X-C motif among AhpD proteins from S. The nitroreductase assay ( Fig. The oxidoreductase AhpF has been found in the prokary- otes Escherichia coil and Salmonella typhimurium (Storz et aL, 1989). hjs oeb laou vlxk mwsahk dcot srpo kaampfel ohqapjk rdqkq mbch mbvd wzxu ffcfo ritsfuu