affinity purification of histidine tagged fnba protein

... of phosphohistidine in proteins and purification of protein -histidine kinases Methods Enzymol 200, 388–414 Kamps, M.P (1991) Determination of phosphoamino acid composition by acid hydrolysis of ... sequence of this enzyme shows no resemblance to any eukaryotic protein kinase The closest homologs to BCKDH kinase are found among the histidine protein kinases of bacteria Direct evidence for histidine ... rapid detection of histidine phosphoproteins in cellular crude extracts Running the phosphorylation reactions in the presence of mM EDTA (instead of Mg2+) exclusively results in histidine phosphorylation...

Ngày tải lên: 24/03/2014, 03:21

... Xa and purification Table summarizes the results of N-terminal sequencing for the major bands of each LASV protein The full length 55-kDa and truncated 46-kDa fragments of LASV Protein Protein ... expression of both LASV glycoproteins, allowing us to achieve the highest yield of recombinant protein per gram of cell mass in an environment appropriate for generation of conformationally correct protein ... resulted in an average protein yield of only ~0.1 mg of MBP-GP1 fusion protein per liter of culture grown in cLB in shake flasks Thus, to obtain a sufficient concentration of MBP-GP1 for our studies,...

Ngày tải lên: 20/06/2014, 01:20

... immunodetection of fractions from asialofetuin affinity chromatography Analysis by Analysis by immunodetection of fractions from asialofetuin affinity chromatography Five mg of the glycoprotein asialofetuin ... 6xHIS RTB p24 Figure of recombinant proteins expressed in E coli and immunodetection Purification Purification of recombinant proteins expressed in E coli and immunodetection Purification was performed ... demonstrate the integrity of the protein after purification (Figure 3, panel C) Purification of E coli expressed p24 HIV and chimeric RTB/p24 proteins In order to improve solubilization of the E coli-producedRTB/p24...

Ngày tải lên: 12/08/2014, 04:21

... ISOLATION OF AN OSMOTIN-LIKE PROTEIN GENE FROM STRAWBERRY AND ANALYSIS OF THE RESPONSE OF THIS GENE TO ABIOTIC STRESSES………………………………………………… 54 EXPRESSION OF A STRAWBERRY OSMOTIN-LIKE PROTEIN GENE, ... contain seven protein complexes, which consist of various combinations of a chitinase, two glucanase-like proteins (GLPs) and a thaumatin-like protein (TLP) Western blot analysis of the strawberry ... substrate of chitinases, chitin, is also a major structural component of fungal cell walls The PR-5 family consists of thaumatin-like proteins and osmotin-like proteins Other PR families include proteinase...

Ngày tải lên: 13/11/2014, 09:25

... binding factor Sp1 Affinity Purification/ Tandem MS/MS - - Affinity Purification/ Tandem MS/MS (Wang et al., 2006) - - Affinity Purification/ Tandem MS/MS (Wang et al., 2006) - - Affinity Purification/ Tandem ... University of Singapore, 2008 1.4 Finding protein- protein interactions Protein- protein interactions form a fundamental level of communication between the many proteins at a cellular level The level of ... suitable for the discovery of protein- protein interactions from less abundant proteins such as transcription factors Epitope tagged proteins generated by the knock-in of tags to the endogenous...

Ngày tải lên: 11/09/2015, 09:05

... calculations of the purification factor, we first calculated ratios of the viral proteins compared to the 88 kD and 44 kD cellular proteins in dpi medium and in purified particles in fractions 11–15 of ... dividing the intensity of the individual viral proteins in fractions 11–15 with the intensity of the proteins in the dpi medium We found that the recovery of the viral proteins was about 5% (Table ... other on basis of the number of living cells at the end of collection We used a set of antibodies directed towards cellular proteins, including cytoplasmic, cytoskeletal and surface proteins We...

Ngày tải lên: 18/06/2014, 18:20

... calculations of the purification factor, we first calculated ratios of the viral proteins compared to the 88 kD and 44 kD cellular proteins in dpi medium and in purified particles in fractions 11–15 of ... dividing the intensity of the individual viral proteins in fractions 11–15 with the intensity of the proteins in the dpi medium We found that the recovery of the viral proteins was about 5% (Table ... other on basis of the number of living cells at the end of collection We used a set of antibodies directed towards cellular proteins, including cytoplasmic, cytoskeletal and surface proteins We...

Ngày tải lên: 20/06/2014, 01:20

... considerable amount of non-specific proteins eluted in conjunction with this protein Final purity of the purified proteins is without any doubt an excellent measure of the performance of protein purification ... Comparison of purification yield and protein recovery of three Nickel-magnetic matrices for His-ProT and His-Mre11 recombinant proteins Purification yield was defined as the sum of the percents of the ... for purification of two Histagged recombinant proteins, His-ProT and His-Mre11, overexpressed in Escherichia coli Results Relative expression of His -tagged recombinant proteins Recombinant proteins...

Ngày tải lên: 11/08/2014, 00:23

... fragment was about 1840 bp 3.2 Expression and purification of polyhistidine -tagged VP2 of PPV We have previously shown that formation of chimeric VLPs of PPV fusion constructs is feasible in pET-PPV ... recombinant protein products purified by IMAC as described below Page of 2.6 Hemagglutination assay Following the method of Senda et al [20], two-fold dilutions of samples of VP2 protein were ... baculoviruses encoding the structural proteins VP2 of PPV N-terminally fused to a polyhistidine tag (6 × his) were engineered to simplify the overall purification process of these the viral antigens The...

Ngày tải lên: 11/08/2014, 21:21

... Isotope-Coded Affinity Tags (ICAT) analysis on global protein profiling of Ng expressed mouse neuroblastoma N2 A cells (N A-Ng) versus N2 A control showed 40% of the downregulated proteins are ... of Ng is Ser36 which was determined by automatic sequencing of major radioactive tryptic peptide after trypsin digestion of the phosphorylated protein In addition, none of the Ser36 mutants of ... exhibited a higher affinity for CaM than wild type in the absence of Ca2+ but a similar affinity in the presence of Ca2+; F36W showed a higher affinity to CaM in the absence or presence of Ca2+ whereas...

Ngày tải lên: 05/10/2015, 22:32

... Isotope-Coded Affinity Tags (ICAT) analysis on global protein profiling of Ng expressed mouse neuroblastoma N2 A cells (N A-Ng) versus N2 A control showed 40% of the downregulated proteins are ... of Ng is Ser36 which was determined by automatic sequencing of major radioactive tryptic peptide after trypsin digestion of the phosphorylated protein In addition, none of the Ser36 mutants of ... exhibited a higher affinity for CaM than wild type in the absence of Ca2+ but a similar affinity in the presence of Ca2+; F36W showed a higher affinity to CaM in the absence or presence of Ca2+ whereas...

Ngày tải lên: 06/10/2015, 20:35

... Schematic diagram of the hexa -histidine tagged cHSPA6 fusion protein 2.2 2.2.1 Optimization of cHSPA6 overexpression in E Coli Effect of temperature on the overexpression of cHSPA6 Figure showed that ... express of recombinant cHSPA6 vector which contain hexahistidine tagged cHSPA6 fusion protein, T7 promoter and by highly specific TEV protease site labeled as X-site Fingure1 Schematic diagram of ... eluted protein fractions were analyzed for protein content on 12% SDS–PAGE 1.6 Silver staining To analyze the purity of pooled protein fractions eluted from gel exclusion chromatography, 25 ng protein...

Ngày tải lên: 24/10/2016, 14:24

... [7] The number of mature liver schizonts increased in the presence of mAbs anti-CSP of P yoelii and the authors suggested that enhancement of infection could be a consequence of an interaction ... receptors for region I of the circumsporozoite protein of Plasmodium falciparum Mol Biochem Parasitol 1997; 90: 33-41 18 Thathy V, Fujioka H, Gantt S, et al Levels of circumsporozoite protein in the ... was 3.07 +/- 0.73 (mean +/- SD)] Efficacy of invasion was calculated as the percentage of ring infected erythrocytes of test cultures over the percentage of ring infected erythrocytes in control...

Ngày tải lên: 02/11/2012, 10:14

... specificity of G-quadruplex recognition depends on the guanidinium group of the Arg in the RGG domain in the C-terminal of EWS Identification of Ewing’s sarcoma protein Fig Affinity of EWS for binding ... 250 0.8 0.6 0.4 0.2 220 320 Fig CD of Htelo in the presence of various amounts of RGG3 Titration of Htelo with RGG3 (1.2, 1.0, 0.8, 0.6, 0.4, 0.2, 0.1 and equiv of RGG3) in 100 mM KCl and 50 mM ... using imagequant software The data were plotted as u (1 fraction of free DNA) versus the protein concentration to determine the Kd, which is equal to the protein at which half of the free DNA is...

Ngày tải lên: 15/02/2014, 01:20

... localize PUF proteins Further pfam analyses using a cut-off E-value of 1.0 over the blast-p output identified 25 proteins containing PUF repeats, which is the largest number of putative PUF proteins ... [24,25,33–36] A B Fig CLUSTAL W alignment of the APUM proteins with the PUF domains most similar to Drosophila PUF domain (A) APUM proteins of group I (B) APUM proteins of group II FEBS Journal 276 (2009) ... mRNA (Fig 3B) [36] We used the APUM-2 protein as a representative member of group I proteins and APUM-7 as a representative of group II APUM proteins (Table 3) Protein RNA interactions was evaluated...

Ngày tải lên: 18/02/2014, 06:20

... of the 4p orbital with the 3d orbital of the metal cofactor Two important pieces of information can be gleaned from the pre-edge peak First, occurrence of this transition implies distortion of ... experiments Protein- bound iron FereneS was added in a final concentration of 20 mm to the protein sample to give a total volume of 600 lL and a protein concentration in the range 5–10 lm The formation of ... superoxide in the course of the reaction The principle of the assay is that the presence of a SOD inhibits the reduction of cytochrome c This was validated by adding a total of 2.5 U of commercial SOD...

Ngày tải lên: 18/02/2014, 06:20

... region of VpU, as indicated by a small number of observed long-range NOEs [24] NMR structure of micelle-associated HIV-1 VpUcyt Another important aspect is the topology of the cytoplasmic domain of ... the expected number of cross-peaks of VpUcyt were obtained with perdeuterated DPC (DPC-d38) at 30 °C A series of HSQC spectra of mm VpUcyt was recorded with varying amounts of DPC-d38 in the sample ... mark the theoretical average values of the downfield shift of 13Ca (top) and 13CO (bottom) as well as of the upfield shift of 1Ha resonances (middle) characteristic of a 100% helical secondary structure...

Ngày tải lên: 18/02/2014, 06:20

... phosphorylation, even though protein synthesis was inhibited [3,20] Thus, it has been suggested that phosphorylation of eIF2a may not be the dominant mechanism of inhibition of translation of normal cellular ... stability by binding to PABP1 [7] In light of the vital roles of PABP1 in mRNA metabolism, it is critical to control the expression of PABP1 Expression of PABP1 is regulated primarily at the translational ... 5¢-UTR of PABP1 mRNA [13,14] The ARS regulates the expression of PABP1 by a negative feedback mechanism In the presence of excess PABP1 in mammalian cells, PABP1 binds to the ARS element of its...

Ngày tải lên: 18/02/2014, 12:20

... containing different domains of the protein, to determine the calcium-binding domain of the protein (A) Schematic drawing of the recombinant proteins Six GST-fused recombinant proteins containing the ... six fusions of GST with recombinant proteins of OMM-64 (rOMM-64) were produced and applied to a 45Ca overlay assay (Fig 3) Of these recombinant Fig 45Ca overlay analysis of fusions of GST and ... be aggregates of proteins and polysaccharides, and may play important roles in formation of the phases and ⁄ or morphologies of the crystals because they consist of acidic glycoproteins and may...

Ngày tải lên: 18/02/2014, 17:20

... homology, the p73 protein can recognize the same response elements as the p53 protein and activate an analogous set of downstream genes Multiple splice isoforms of the p73 protein have been found ... PGM4 targets) Effects of DNA cis-platination on sequencespecific DNA binding of p73 proteins We tested the influence of DNA modification with cisplatin on the binding of two p73 isoforms, p73d and p73b, ... destabilization of the p53–DNA interaction (or even prevention of target recognition by the protein) DNA binding of the C-terminally truncated p53(1–363) protein Fig The effects of DNA modification...

Ngày tải lên: 19/02/2014, 05:20