expression purification and biophysical studies of chromodomain proteins

... diagnostics and epidemiologic surveys Methods Strains, plasmids, and enzymes Expression and purification of LSV coat protein Figure Expression and purification of LSV coat protein A, Expression and purification ... 23th candidate proteins had scores from 82 to 178, those were all coat proteins of various isolates of LSV, and protein scores greater than 67 were significant (p < 0.05) 15 of 28 mass values of ... Alignment of the amino acid sequence of coat protein of different LSV isolates and other carlavirus Figure Alignment of the amino acid sequence of coat protein of different LSV isolates and other...

Ngày tải lên: 12/08/2014, 04:21

... Typical lengths of genomic DNA in eukaryotic and prokaryotic cells are in the ! ! magnitude of meters and millimeters, respectively They correspond to random coiled sizes of 3"105 nm and 104 nm, ... the expression of diverse virulence genes The Nterminus of SsrB contains the site of phosphorylation and the C-terminus consists of a DNA-binding domain Dimerization occurs upon DNA binding and ... sugarphosphates The two strands rotate about the same axis and form two grooves with the large groove of 22 Å and small groove of 12 Å Nucleobases are connected with the sugar backbones and they form pairs...

Ngày tải lên: 09/09/2015, 11:25

... domain is mainly built up by residues from the C-terminus and consists of a two-stranded anti-parallel b-sheet composed of b2 and b9 and an a-helix from the C-terminus (Fig 1A) The two subunits ... side chain of Lys173 interacts with both O6 and N7 (Fig 1C) The 2¢OH of the ribose is hydrogen bonded to the main chain and the side chain of Asp142 (Fig 1C) Both hydroxyl groups of the ribose ... PRTFDC1 towards hypoxanthine and guanine is only 0.26% and 0.09%, respectively, of the activity of HPRT, the role of the PRTFDC1 in purine metabolism remains unclear and it is uncertain whether...

Ngày tải lên: 15/02/2014, 01:20

... A-chain and B-chain of nPS, and in the case of His37, the v1 torsion angle changes from 63.34° to )67.80° and )52.11° for the A-chain and B-chain of nPS, respectively The imidazole ring of His34 ... we report the results of the structural studies of the pantoate-bound form of the N-terminal domain of E coli PS (nPS) The structure and substrate-binding interactions of E coli nPS have been ... specific residues of nPS as a function of substrate(s) The resonances of free nPS are shown in red, those of nPS bound to D-pantoate are shown in blue, and those of nPS bound to D-pantoate and ATP are...

Ngày tải lên: 16/02/2014, 09:20

... shown) Cloning of anserinase cDNA PCR was performed using a set of primers (A and B) and the first-strand cDNA for 3¢ rapid amplification of cDNA ends (RACE), prepared from total RNA of 6004 Nile ... including 56 bp of 5¢ UTR, 1482 bp of ORF, and 299 bp of 3¢ UTR The coding region of the sequence was translated into 494 amino acids, which included a typical signal peptide of 18 amino acids and two ... activity of anserinase was 6006 detected strongly in kidney, brain, liver and ocular fluid, and weakly in skeletal muscle and spleen of Nile tilapia The mRNA expression in the brain and kidney...

Ngày tải lên: 19/02/2014, 07:20

... possible dimeric structures of LPL, the head-tohead and the head-to-tail models, the studies of the chimeric proteins of hepatic lipase and LPL and the tandem repeat approach of LPL [21,34] support ... of hydrogen, modification of bonds, potentials of forcefield, and fixation of heavy atom, backbone, and Ca, were performed for molecular mechanics calculations and then the energy minimization of ... (black) and two distinct structures of the lid (orange) Ó FEBS 2002 Dimeric model structure and function of lipoprotein lipase (Eur J Biochem 269) 4705 Table Heparin affinity of C-terminal domain of...

Ngày tải lên: 21/02/2014, 15:20

... NMR studies [16,19,20,23] The energy of folding these various forms and the energetic cost of converting one form to another is of fundamental importance for understanding telomere biology and ... understanding telomere biology and the interactions of small molecules and proteins with telomeric DNA The folding and unfolding of quadruplexes may be of profound physiological importance because quadruplex ... practical details of UV melting studies of G-quadruplexes, with an thorough discussion of common problems and pitfalls [32] The use of fluorescence to monitor the thermal denaturation of quadruplex-forming...

Ngày tải lên: 06/03/2014, 09:22

... calculated from the ratio of the rate constants kon and koff as follows: Ka ẳ kon koff 2ị Association constants (Ka) of CRP with the three investigated sequences of DNA lacF, galF and ICAPF are summarized ... ẳ 0.19 high afnity and sequence specicity [2] A variety of biochemical and biophysical studies [1316], including our fast-kinetics studies [17,18], as well as steady-state and time-resolved uorescence ... Purication of proteins The isolation and purication procedures of wild-type CRP and CRPW85A mutant were carried out in the same manner as previously described [7] The purity of the proteins was...

Ngày tải lên: 07/03/2014, 16:20

... Rao and Dr J Josse for critical review of the manuscript, and Professor W Loffelhardt (University of Vienna, Austria) for ¨ his assistance in N-terminal protein sequencing and some MALDI-TOF MS ... molecular mass of 19 216 Da and a pI of 4.69, in good agreement with MALDI-TOF MS (Fig 1A) and chromatofocusing data, respectively The other three N-terminal sequences determined in this work, of Anabaena ... UÆmg)1 protein and recovery levels of 20– 30% In all cases, the analysis by SDS ⁄ PAGE and native PAGE of purified preparations showed only one protein band of 20–22 kDa (Figs and 2; supplementary...

Ngày tải lên: 16/03/2014, 11:20

... future analysis Purification of muscle proteins Experimental procedures Construction of expression plasmids and site-directed mutagenesis The pPIC9 expression vector and P pastoris strain GS115 ... demonstrated by the biophysical assays Our studies have provided additional contributions to the understanding of the effects of these mutations on the clinical symptoms of patients carrying ... microcalorimetric study of Tm denaturation was performed using a scanning microcalorimeter MicroCal Ultrasensitive VP-DSC and standard software for data acquisition and analysis Tm concentrations were of 15 lM...

Ngày tải lên: 16/03/2014, 18:20

... turns) and 16% of other contributions for both the native and refolded forms, with a rmsd of 1% The deconvolution of Mfrutalin showed 13% of a helix and 12% of b components (antiparallel and parallel ... where N and U represent the native and reversibly unfolded forms of the frutalin and kN and kU, the equilibrium constants of the unfolding transitions from the N to the U state The fraction of unfolded ... Nfrutalin and Rfrutalin-Ch were deconvoluted by convex constraint analysis [19,20] as described in Materials and methods, and showed 86% of beta components (antiparallel and parallel b sheet, and b...

Ngày tải lên: 17/03/2014, 23:20

... In IB-367 and BM-1, the presence of strong daN(i,i + 1) and weak (or absence of) dMN(i,i + 1) and dbN(i,i + 1) NOE connectivities in the Gly3–Arg8 and Cys14– Gly16 regions of IB-367 and in the ... Arg1 and Arg17, and in the turn in the presence of Arg8 and Arg10 (see Figs and 4) Our conformational studies showed that BM-1 formed a twisted b-sheet structure, similar to that of native PG-1 and ... activity of the peptides A better understanding of the mode of action of these peptides is crucial for the development of a new generation of antibiotics It is known that, in the absence of both...

Ngày tải lên: 22/03/2014, 21:20

... Lane and 9, respectively, P and S fractions of 0.5% Triton X-100 plus 0.32 M NaCl treatment of thrombin digested microsomes Lane 10 and 11, respectively, P and S fractions of M NaCl treatment of ... respectively Lanes and 5, respectively, P and S fractions of untreated thrombin digested microsomes Lanes and 7, respectively, P and S fractions of 0.5% Triton X-100 treatment of thrombin digested ... mutational studies of chalcone synthase have demonstrated that decarboxylation of malonyl-CoA is independent of acylation of the active site cysteine [24] In these studies, substitution of the active-site...

Ngày tải lên: 24/03/2014, 03:21

... concentration of XAIP (b) and with lM concentration of chitinase enzyme (c) for h, and (B) in the absence of any protein (a), with 100 lM concentration of XAIP (b) and with 100 lM concentration of chitinase ... internal space of the TIM barrel considerably Of the three observed cis peptides, two (Gly33–Phe34 and Trp253–Asp254) are found at the ends of b-strands b2 and b8, respectively These are part of the ... structure of XIP-I Recently, the structure of XIP-I has been reported [14] It binds to two types of xylanase from the subgroup of family 18C proteins: GH10 and GH11 xylanases The overall scaffolding of...

Ngày tải lên: 29/03/2014, 09:20

... mechanism of the AspB-catalyzed reaction and pKa values of potential acid and base catalysts by pH-rate studies From the ascending limb of the pH-rate prole of AspB, a residue with a pKa of 7.1 ... obtained from grafit software Kinetics of reversible inhibition of AspB, T141A and H188A by 3-nitropropionate and D-malate The reversible inhibition of AspB and the T141A and H188A mutants was ... pH-rate and inhibition studies to further illuminate the mechanistic role of certain residues The results obtained in these studies are interpreted and related to the mechanisms of AspA and other...

Ngày tải lên: 29/03/2014, 23:20

... changes in expression of thousands of genes [9,12,13] Another challenge is that the specificity and sensitivity of a given RNA profiling platform are affected by the abundance and variability of the ... spectrophotometry and quality was assessed using an Agilent Bioanalyzer and the Agilent RNA 6000 Nano kit (data not shown) Gene expression profiling Aliquots of each sample were profiled for gene expression ... detection and discrimination of low -expression genes by increasing expression signal across a wider range of low-intensity probes Another important characteristic of cDNA targets is the reduction of...

Ngày tải lên: 18/06/2014, 16:20

... reports as well [31] The XPS data of Zn2p and Mn2p further provide evidences for the incorporation of Mn2+ ions into ZnO To obtain a better understanding of the formation of Mn-doped ZnO HMSs, a probable ... coordination route of EG which directs the formation of Mn-doped ZnO HMSs in view of morphology transformation A lot of EG molecules are adsorbed on the (001) surface of the ZnO crystals and block the ... spectra The origin of the peaks at 425 and 475 nm could be ascribed due to the transition occurring from Zn interstitials to the valence band, and the peak of 425 nm may be the result of the singly...

Ngày tải lên: 20/06/2014, 20:20

... presence of two absorption bands at 1273.7 and 3421 cm−1 belonging to the C=S and NH groups, respectively, and by absence of SH Density functional theory (DFT) studies DFT calculations of the IMHC ... nature of functional group of IMHC The appearance of a broad strong band in the IR spectra of IMHC in 3421 cm–1 is assigned to N–H stretching vibrations of the primary amine group The spectrum of ... synthesis of some bioactive compounds such as ß-lactams [16–18], and employed as ligands for the complexation of metal ions [19] SBs and their complexes are largely studied because they interested and...

Ngày tải lên: 20/06/2014, 20:20

... the design of the study and performed the electrical studies SK and MA carried out the structural studies SAA and MSA performed the thermal studies MAMK and MA also involved in writing of the manuscript ... observed The copper and the carbon peaks correspond to the carbon coated copper grid of TEM No peaks of other impurity have been detected Therefore, the EDX profile of sample (inset of Figure 2) indicates ... http://www.nanoscalereslett.com/content/6/1/434 Page of A B Figure FETEM and HRTEM images of silver nanoparticles (a) FETEM image of silver nanoparticles (b) High-resolution image of a single silver nanoparticle and inset shows...

Ngày tải lên: 21/06/2014, 02:20

Ngày tải lên: 21/06/2014, 02:20