synthesis purification and biochemical characterization
Báo cáo y học: "HIV-1 Tat protein enhances Microtubule polymerization" docx
... should have a tremendous impact for AIDS therapy Methods Protein synthesis, Purification and Biochemical Characterization Tat variants and their derived peptides were synthesized with an ABI 433A ... polymerization, quantification of microtubules and electron microscopy directed by VP GRC, SL, SO, DE and JW synthezised the peptides and characterized them biochemically and biophysically CP performed FACS ... band intensity of pellets (P) are enhanced and the band intensity of supernatants (S) are decreased compared with the control tubulin alone Increasing the concentration of Tat to µM (Tat 8) and...
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Tài liệu Báo cáo khoa học: Tissue expression and biochemical characterization of human 2-amino 3-carboxymuconate 6-semialdehyde decarboxylase, a key enzyme in tryptophan catabolism pptx
... curves and data were normalized to the copy number of 18S RNA Expression of ACMSD isoforms in E coli pGEM-ACMSD I and pGEM-ACMSD II were digested with XhoI and Bpu1102I and cloned into pET-15b and ... swiss-pdbviewer and visual molecular dynamics [49] Acknowledgements We thank G Magni and S Ruggieri for helpful comments and suggestions and T Begley (Department of Chemistry and Chemical Biology, ... Escherichia coli and Pichia pastoris ACMSD I and II cDNAs were subcloned into pET15b and pET32b E coli expression vectors, providing the recombinant proteins with a N-terminal His6-tag, and a thioredoxin-tag,...
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Tài liệu Báo cáo khoa học: Structural and biochemical characterization of a human adenovirus 2/12 penton base chimera pptx
... insertion of $ 230 residues (residues 133–367) between strands D and E and a smaller insertion of $ 50 residues (residues 404–458) between strands F and G The hypervariable loop, FEBS Journal 273 (2006) ... expression, the SF21 and Hi5 cell lines and the Bac-to-Bac expression system and vectors were from Invitrogen (Carlsbad, CA, USA) Protease inhibitors were from Roche (Basel, Switzerland) and Ni-NTA resin ... penton base (yellow), hAd2 ⁄ 12 (blue), and hAd2 fiber peptide complex (pink) The fiber peptide is drawn as ball -and- stick and colored by atom fiber peptide bound and unbound structures reveal a high...
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Tài liệu Báo cáo khoa học: Molecular and biochemical characterization ofD-phosphoglycerate dehydrogenase fromEntamoeba histolytica A unique enteric protozoan parasite that possesses both phosphorylated and nonphosphorylated serine metabolic pathways docx
... a limited set of enzymes involved in amino acid synthesis of Ser from Gly and Ala from Cys and conversions between Asp and Asn and between Glu and Gln [49] Serine metabolic pathways are often ... cyanobacteria, Clostridiales, Actinomycetales and archaebacteria belong to Type I, while b- and c-proteobacteria and Bacteroidales belong to Type IIA and Type III, respectively It is worth noting ... (from archaea and bacteria) [43,44] and for GDH (from e-proteobacteria) [30], or, in contrast, that Bacteroidales obtained the gene from E histolytica or E caudatum Purification and characterization...
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Tài liệu Báo cáo Y học: Structural and biochemical characterization of neuronal calretinin domain I– II (residues 1– 100) Comparison to homologous calbindin D28k domain I–II (residues 1 –93) pdf
... for both CR and Calb [22,24,30] The biochemical properties of CR and Calb are similar to those of calmodulin, the best-known EF-hand calcium sensor It is difficult to localize the biochemical ... us to analyze the biochemical and structural properties of CR I– II and to compare them with Calb I–II Our data show that CR I –II and Calb I –II share virtually the same EF-hand structures However, ... underlined An EF-hand consensus sequence is placed between the aligned EF-hand I and EF-hand II sequences [3] The helices are shown as boxes with conserved hydrophobic (h) and other amino acids...
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Báo cáo Y học: Purification and biochemical characterization of some of the properties of recombinant human kynureninase pptx
... plotted using the CRICKETGRAPH and GraphPad PRISM3 software packages, and the kinetic parameters Km and Vmax were obtained using non-linear regression Lineweaver–Burk and Dixon [14] plots were used ... reducing agents SDS and mercaptoethanol to determine the native dimeric molecular mass resulted in the appearance of two bands at 52.5 and 95 kDa (gel image not shown), and this shows that ... spectrofluorimetrically at 37 °C, with excitation of the product 3-hydroxyanthranilate at 330 nm and emission at 410 nm and 310 nm and 417 nm respectively for anthranilate, by the method of Shetty & Gaertner [7]...
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Báo cáo Y học: Structural and biochemical characterization of calhepatin, an S100-like calcium-binding protein from the liver of lungfish (Lepidosiren paradoxa) docx
... crucial for dimerization and monomer stability, in Sl00 A4 and other S100 proteins [23] (Fig 3), are clustered between helix I and IV (Fig 5B), in the same way as in Sl00 A4 and other S100 proteins ... the crystal and/ or NMR structures of other members of the family The molecule is depicted in strand representation, and a helices are numbered from I to IV (B) Residues Leu7, Arg8 and Phe11 from ... a-helix I, and Trp60, Phe63, Ala66 and Phe67 from a helix IV (located at equivalent positions to those crucial for S100 A4 dimerization and monomer stability [23]) are shown in dark and light...
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Báo cáo khóa học: Identification of a gene encoding Lon protease from Brevibacillus thermoruber WR-249 and biochemical characterization of its thermostable recombinant enzyme pptx
... used as the standards and their elution volumes, Ve, were determined The standard curve was plotted with the logarithm of molecular mass against Ve/V0 of the standard protein Peptidase and ATPase ... phosphatase Materials and methods Bacterial identification and culture conditions DNA manipulation and sequence analysis Plasmid DNA preparation, purification of DNA from agarose gel, and restriction ... was extracted from Br thermoruber by standard methods [23] and digested with HindIII and SalI The digested DNA fragments were ligated with cassette adaptors and then used as a template for the following...
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Báo cáo khoa học: An a-proteobacterial type malate dehydrogenase may complement LDH function in Plasmodium falciparum Cloning and biochemical characterization of the enzyme potx
... manufacturer’s protocol Cloning of Pf MDH, Pf LDH and Pf MQO Pf MDH was cloned for functional and biochemical characterization The other two genes Pf LDH and Pf MQO were also cloned to obtain the DNA ... evaluation of Vmax and kcat values for malate/NAD and OAA/NADH (Table 1) The saturating concentrations of OAA and NADH were found to be 250 lM and 150 lM, respectively, while for malate and NAD these ... pathways i.e nonmevalonate pathway of isoprenoids biosynthesis, fatty acid biosynthesis, purine salvage, pyrimidine biosynthesis, shikimate pathway and synthesis of GPI anchors [4] The parasite, however,...
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Báo cáo khoa học: Identification and biochemical characterization of the Anopheles gambiae 3-hydroxykynurenine transaminase pot
... identification, the bacterial heterologous expression and the biochemical characterization of 3-HKT from A gambiae These results represent the first biochemical report concerning an enzyme 5654 F Rossi ... a better understanding of the XA metabolism in the mosquito and offers a new possible target for the development of insecticides and ⁄ or transmission blocking agents Results and Discussion Isolation ... Within such a scenario, the isolation and biochemical characterization of recombinant A gambiae 3-HKT that we report here, represents a step forward into the understanding of XA metabolism in FEBS...
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Báo cáo Y học: Agmatine oxidation by copper amine oxidase Biosynthesis and biochemical characterization of N-amidino-2-hydroxypyrrolidine pdf
... 10)3 >5 · 10)2 c >10)2 e c pH 7.5 and 37.0 °C Present study b pH 7.8 and 37.0 °C From [8] c pH 7.5 and 37.0 °C From [16] d pH 6.8 and 21.0 °C Present study pH 7.0 and 25.0 °C From [30] 890 P Ascenzi ... rate-limiting catalytic constant (kcat) and of the Michaelis constant, as determined in the absence and presence of the inhibitor (K and K app , m m respectively), for NOS-I and NOS-II catalyzed conversion ... 1.0 · 10)5 M and 1.0 · 10)3 M, and the N-amidino-2-hydroxypyrrolidine concentration ranged between 2.0 · 10)5 M and 8.0 · 10)5 M The enzyme activity was linear up to 10 of incubation and results...
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Báo cáo khóa học: Isolation and biochemical characterization of two soluble iron(III) reductases from Paracoccus denitrificans docx
... values of Km and kcat are: (0.4 ± 0.2) mM and (0.91 ± 0.02) s)1 for Fe(III)EGTA (1.6 ± 0.8) mM and (0.16 ± 0.03) s)1 for Fe(III)EDTA (0.4 ± 0.2) mM and (0.08 ± 0.01) s)1 for Fe(III)citrate, and (0.4 ... chromophore uniquely attributable to this enzyme On the other hand, the solution of FerB was yellowish in color and exhibited absorption bands at 380 and 448 nm (Fig 3), diagnostic for the presence of a ... FerA and to a ping-pong mechanism for FerB Non-linear regression values of kcat are (14 ± 1) s)1 and (28 ± 6) s)1 for the Fe(III) reductase and FMN reductase activity of FerA, respectively, and...
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Identification and biochemical characterization of tetrahydrolipstatin targets in m bovis BCG at different metabolic states
... infect and survive in the host One of the highly conserved strategies is to modify the host response and/ or utilize host factors for survival and colonization Thus, identification and characterization ... bacilli membrane and culture filtrate fractions 148 6.5.2 PTM-mediated protein regulation 149 6.5.3 Elucidating the substrate of lipH 149 6.5.4 Validation and characterization of BCG2654c and lipM 150 ... occurs earlier, and at not pose and disease progression is complex and highly lower bacterial load In most animal models, tenuating strains of Mtb through the deletion dynamic and appears to be...
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Biochemical characterization of thermophilic lignocellulose degrading enzymes and their potential for biomass bioprocessing
... (filer paper), BWX and PWSD The enzyme kinetic studies for cellulase and xylanase (Km and Vmax) were performed with 1-10 mg/ml of CMC and BWX, respectively [35] The crude cellulase and xylanase were ... albumin as standard [38] All experiments were run in duplicate and standard deviations (SD) were calculated using Microsoft Excel and results were presented as average ± SD Results and discussion ... Amaral-Collar M.T Characterization of a thermotolerant and alkalotolerant xylanase from a Bacillus sp Appl Biochem Biotechnol 1998, 73, 159-172 [53] Huang X.P., Monk C Purification and characterization...
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Tài liệu Báo cáo khoa học: Pyrimidine-specific ribonucleoside hydrolase from the archaeon Sulfolobus solfataricus – biochemical characterization and homology modeling doc
... hypoxantine, guanosine and guanine, uridine and uracil, cytidine and cytosine were 12.2 and 6.2 min, 10.5 and 4.7 min, 15.2 and 6.1 min, 6.8 and 4.2 and 6.6 and 3.9 respectively The amount of purine ... employed and the elution was carried out with : 94 (v v) mixture of 95% methanol and 0.1% triuoroacetic acid in H2O The retention times of adenosine and adenine, inosine and hypoxantine, guanosine and ... A, we were able to calculate the volume of buried and surface cavities for SsCUNH and the template, both in the monomer and in the tetramer, and we found that the volume of buried cavities found...
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Tài liệu Báo cáo khoa học: Biochemical characterization and inhibitor discovery of shikimate dehydrogenase from Helicobacter pylori docx
... H pylori shikimate dehydrogenase (HpSDH) was cloned, expressed, and purified in E coli system, and its biochemical and enzymatic characterizations were also carried out Furthermore, by using the ... (Eqn 2) On the other hand, compounds 1, 4, and are noncompetitive inhibitors, and compounds and are competitive inhibitors with respect to NADP Table summarizes the IC50 values and kinetic inhibition ... 15.4 In conclusion, we have firstly cloned and expressed HpSDH enzyme, and the biochemical characterization of HpSDH is expected to favor better understanding the SDH features Moreover, by high...
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Tài liệu Báo cáo khoa học: Biochemical characterization of human 3-methylglutaconyl-CoA hydratase and its role in leucine metabolism docx
... seems, that the fronting, the shoulder and the tailing is due to acetyl-CoA and not due to any other compound (B) The peaks 3, 4, and were isolated by HPLC and used as substrates for AUH The AUH ... its synthesis and developing a new assay In addition, a mutant form of AUH (A240V) derived from an MGA1 patient was tested using 3-MG-CoA Results Overexpression of AUH in Escherichia coli and ... protein and it was thus assumed that the mature form of human AUH in brain has a molecular weight of 32 kDa (AUHp32) [15] For the kinetic characterization of AUH described in the work at hand, AUH...
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Tài liệu Báo cáo khoa học: Biochemical characterization of Bacillus subtilis type II isopentenyl diphosphate isomerase, and phylogenetic distribution of isoprenoid biosynthesis pathways doc
... template and the oligonucleotides 5¢-TTGGTGGGATCCGTGACTCG AGCAGAACGAAAAAGAC-3¢ and 5¢-GGCTTTGTCG ACTTATCGCACACTATAGCTTGATG-3¢ as primers (restriction sites are underlined and start- and stop-codons ... XL1-Blue (Stratagene [32]) and M15 (pREP4) [33] affording the recombinant strains XL1-pQEidi2 and M15-pQEidi2 Preparation of the recombinant Idi-2 protein Fig Biosynthesis of IPP and DMAPP 1-hydroxy-2-methyl-2-(E)-butenyl ... FMN, mM sodium acetate, and 10.8 mM IPP and 10.8 mM DMAPP, respectively; *, internal standard (acetate) FMN, we observed the appearance of the signals of both methyl groups and of the methine group...
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Tài liệu Báo cáo khoa học: Biochemical and molecular characterization of hazelnut (Corylus avellana) seed lipoxygenases pdf
... C6 and C9 aldehydes (and the corresponding alcohol) were detected in all the samples and, similarly to the pattern of LOX expression and activity, their levels were higher in stage I seed and ... the shelf life and the organoleptic features of hazelnut seeds, we carried out a study to characterize hazelnut lipoxygenases at the biochemical and molecular level Materials and methods Plant ... start and the stop codon The introns and exons are drawn in proportion to their length (B) Nucleotide and deduced amino acidic sequences of the hazelnut LOX gene The inverted repeats are in bold and...
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Báo cáo khoa học: Biochemical characterization of USP7 reveals post-translational modification sites and structural requirements for substrate processing and subcellular localization pptx
... Coomassie stained and major bands were excised and eluted in order to perform N-terminal sequencing Tandem affinity purification and mass spectrometric analysis of USP7 N-terminal and C-terminal TAP-tagged ... Fernandez-Montalvan et al Biochemical characterization of USP7 Fig Enzymatic characterization of USP7 (A) Progress curves for the USP7-catalyzed hydrolysis of Ub-AMC (j), SUMO-1-AMC (d) and Nedd8-AMC ... expression vectors and USP7 cDNA as well as Shirley Gil-Parrado, Bruno Martoglio, Martin Renatus and Jorg Eder (Novartis, Basel, Switzerland) ¨ ´ for support and helpful discussions A Fernandez´ Montalvan...
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