diverse domains of cytosine 5 dna methyltransferases structural and functional characterizati

... water and resuspended as a 50 % slurry Purified proteins ( 150 pmol) were added to 1 75 lL of binding buffer ( 25 mM Tris/HCl, pH 7 .5, 150 mM NaCl, 2 .5 mM MgCl2, 0.1% Triton-X100) and 25 lL of slurry ... of the deletion of the RNA- and membranebinding domains of S pneumoniae Era on the GTPase activity of the protein To assess the potential effects of removal of the RNA- and membrane-binding domains ... evidence that the RNA- and membranebinding domains are overlapping Materials and methods Materials Fig Functional domains of Streptococcus pneumoniae Era (A) The structural motifs of S pneumoniae Era...

Ngày tải lên: 31/03/2014, 07:20

... sequenced and determined to be 254 3 bp in length The comparison of nucleotide sequences between the gene and the cDNA [8] showed that the fragment contained the 5 half of the gene including part of ... electrophoresed in 5% acrylamide gels excised and eluted by diffusion into 0 .5 M NH4OAc Binding was carried out in 15 µl of 10 mM Tris (pH 8), mM EDTA, 100 mM NaCl, mM DTT, 10% glycerol and µg of denatured ... experiments to 0 .5 µg of poly dI-dC was also included in the mixes At the end of the incubation period 1/10th of the mix volume of loading buffer was added and samples were loaded on a 5% polyacrylamide...

Ngày tải lên: 08/08/2014, 14:20

... Ramachandran plota Iba2t Iba2o 70 .5, 70 .5, 95. 2 71 .5, 186 .5, 51 .4 P3221 0.9184 10 457 P21212 0.9184 38 251 302. 45 (2 .58 2. 45) 302. 15 (2. 252 . 15) 99.6 (99.9) 99.7 (100) 7.9 (8.0) 6.0 (64.8) 20 .5 (3 .5) ... 8.2 ( 75. 3) 16.1 (3.0) 2. 45 (2 .51 2. 45) 896, 40 2. 15 (2.212. 15) 3406, 221 20.4 (34.1) 23.2 (24.9) 36.6 0.016 1.7 95. 0 5. 0 21.6 (26 .5) 25. 7 (32.3) 38.1 0.017 1.6 94.8 5. 2 a PROCHECK [ 45] : most ... value [36] of 4.0 A3ặDa)1 and a solvent content of 69% Iba2o crystals belong to space group P21212 with cell stants a = 71 .5, b = 186 .5 and c = 51 .4 A A VM value Human Iba proteins of 2 .5 A3ặDa)1...

Ngày tải lên: 16/03/2014, 06:20

... goal of this study was to investigate the structural and functional characteristics of smelt type II AFP The effects and inter-relationship of metal ion binding, N-linked glycosylation, and quaternary ... concentrations of 0, 0.16, 0.3, 0.6, 1. 25, 2 .5, 5, 10, and 20 mM, respectively (A) Intact AFP in the presence of CaCl2 (B) Deglycosylated AFP in the presence of CaCl2 (C) Intact AFP in the presence of EDTA ... intensity of a higher-molecular-mass band The apparent molecular mass of this band was 38 kDa, which is substantially higher than the molecular mass of the monomer band, indicating the presence of a...

Ngày tải lên: 24/03/2014, 03:21

... 4.8 68 186 0. 05, 0.11 7.7 25 0.07, 0.08 2 .5 Local C of SV and SE (A280)e ApoE4-(72-166) Sc (Svedberg) Md (kDa) 0.07, 2.7 24 0.16, 0.04 4.7 51 0.18, 0. 05 251 0.03, 0.06 8.6 210 0. 05, 0.12 21 0.08, ... 2.2 22 0.01, Local C of SV and SE (A280)e Local C of SV and SE (A280)e 0.02, 4.9 4.9 52 0. 15, 4.8 51 0.17, 0.07 0.04, 0.09 8.4 232 0.03, 0.06 8 .5 217 0.04, 0.19 1.3 19 0. 35, 0.13 1.2 20 0.34, ... the same species of apoE4-(72166) The second major species was a broad peak at s = 3 .5 to 6 .5, with a total occupancy of 46 % for apoE2-(72-166), 55 % for apoE3-(72-166), and 59 % for apoE4-(72-166)...

Ngày tải lên: 10/08/2014, 05:21

... respectively) were recorded in the laboratory Structural and Functional Assays Cell Viability Assay Structural and functional viability of saphenous vein endothelial and smooth muscle cells were assessed ... identified by XYZ scanning and imaged at depths of 50 μm away from the site of excision in transverse sections of the SV segments Page of Western Blotting Protein extraction and electrophoresis were ... extraction on structural and functional viability of saphenous vein endothelium Journal of Cardiothoracic Surgery 2011 6:82 Submit your next manuscript to BioMed Central and take full advantage of: •...

Ngày tải lên: 10/08/2014, 09:21

... Purification of recombinant Cr-TRP16 43 2.3.2 Overall structure 45 2.3.3 Sequence and structural homology 47 2.3.4 Dimerization of Cr-TRP16 51 2.3 .5 H- 15N-HSQC ... 51 iv 2.3.6 Analytical ultracentrifugation (AUC) 54 2.3.7 Cr-TRP16 increases TNF- induced nuclear translocation of p 65 and p5 057 2.3.8 Cr-TRP16 augments TNF- induced NF-B DNA binding ... protein design and engineering 22 Figure 2.1: FPLC profile of Cr-TRP16 45 Figure 2.2: Dynamic light scattering (DLS) profile of Cr-TRP16 45 Figure 2.3: Structure of Cr-TRP16 ...

Ngày tải lên: 09/09/2015, 18:55

... receptor ligands 49 vii 1.13 Aim and scope of the thesis 50 CHAPTER TWO: ISOLATION AND PURIFICATION OF HADITOXIN 52 2.1 INTRODUCTION 52 2.2 MATERIALS AND METHODS 55 2.2.1 Materials 55 2.2.2 Sequence ... receptors 5. 2.4 Bacterial strains and plasmids 172 5. 2 .5 Cloning of haditoxin gene into pLICC plasmid 174 5. 2.6 Sequence analysis 174 5. 2.7 Recombinant protein expression 1 75 5.2.8 Extraction of the ... Isolation and purification of haditoxin 59 2.3.2 Identification of haditoxin 62 2.3.3 Assessment of homogeneity of haditoxin 62 2.3.4 Secondary structure analysis of haditoxin 65 2.4 DISCUSSION 66 2.5...

Ngày tải lên: 10/09/2015, 08:37

... PCR of successfully ligated and transformed CaM and ΔNav1.6 clones 89 Fig 3.3 12 .5% SDS-PAGE purification profile of His-ΔNav1.6 90 Fig 3.4 Gel filtration profile of His-ΔNav1.6 91 Fig 3 .5 Purification ... Purification and preliminary characterisation of CaM 94 Fig 3.6 12 .5% SDS-PAGE profile of His-ΔNav1.6 trapped by CaM-sepharose 95 Fig 3.7 Superimposed elution profiles of His-ΔNav1.6, Ca2+/CaM and HisΔNav1.6: ... 1.1 List of common and important PTMs 23 Table 1.2 IQ motifs of established and potential CaM target proteins 37 Table 2.1 Data collection and refinement statistics of Cbl-TKB complexes 55 Table...

Ngày tải lên: 11/09/2015, 09:11

... Calculation 52 2 .5. 8 Relaxation Data Analysis 53 2 .5. 9 Docking of Def-A with LPS 54 CHAPTER STRUCTURE AND DYNAMICS OF DEF-A BOUND TO SDS AND ITS BACTERICIDAL ACTIVITY 3.1 Design, expression and purification ... addition (B) Plots of the heat change as a function of POPG:Def-A molar ratio 69 1D 1H Spectra of 0.5mM Def-A In the absence of SDS at 15 °C (A) and in presence of 50 mM SDS micelles at 35 °C (B) 71 ... phagocytin and CAP molecules of rabbit and human were reported in 19 85 (Selsted et al., 1985a and 1985b) and then recognized as natural peptide antibiotics and renamed as defensins (Ganz et al., 19 85) ...

Ngày tải lên: 14/09/2015, 08:37

... Transformation of ligation mix into DH5-α competent cells 51 2.1.7 PCR screening of transformant 51 2.1.8 Isolation of DNA plasmid 52 2.1.9 Plasmid DNA sequencing 2.2 49 53 Protein expression and purification ... development of hypoallergenic mutant 5. 2 Structure characterization of Blo t 153 5. 3 Future direction 154 References 157 Appendix 1 75 viii Summary Dust mite has been one of the major causes of allergic ... experiment 2 .5 Sub-cloning, expression and purification of Blo t 75 2.6 NMR experiments and chemical shifts assignment of Blo t 75 Chapter 3: Structure characterization and IgE epitope mapping of Der...

Ngày tải lên: 14/09/2015, 18:50

... 200 50 50 10 10 50 100 150 200 250 S 200 50 250 50 1200 100 150 [Hx] (µM) 2.0 PRTFDC1 50 100 150 200 250 S HPRT D C 0.0 12 15 S/V V (µmol·min–1·mg–1) 0.3 S/V V (µmol·min–1·mg–1) 0.4 100 150 [Hx] ... triplicate and given as the mean ± SD Ligand DTagg (°C) 50 0 lMa DTagg (°C) 10 mMb PRPP IMP GMP CMP UMP AMP XMP 5. 8 5. 1 4 .5 )0.2 )0.2 0.0 0.6 12.7 7.8 10 .5 0.2 0 .5 1.4 3 .5 Ligand DTagg (°C) 50 0 lMc ... [Hx] (µM) 200 250 50 1.0 40 30 20 0 .5 10 800 600 0.10 0.08 400 S/V V (µmol·min–1·mg–1) 60 S/V Fig Characterization of PRTFDC1 and HPRT with hypoxanthine and guanine Michaelis–Menten and Hanes–Woolf...

Ngày tải lên: 15/02/2014, 01:20

... 350 400 450 λ (nm) 50 0 55 0 600 650 700 λ (nm) 0.06 0.04 350 400 450 50 0 λ (nm) 55 0 0. 15 0.10 0. 05 0.02 0.00 300 0.20 600 650 0.00 300 700 400 450 50 0 λ (nm) 55 0 600 650 700 0.09 ~56 0 58 0 nm 0.12 ... properties of M tuberculosis WhiB proteins 0.08 0. 05 0.030 0.01 400 450 55 0 600 650 0.00 300 700 340 nm 400 450 50 0 55 0 600 650 700 0.18 WhiB3, 50 µM WhiB3, 50 µM, alkylated 0.16 WhiB4, 50 µM WhiB4, 50 ... 50 µM WhiB2, 50 µM, alkylated 337 nm 420 nm 0.07 ~56 0 58 0 nm Absorbance 0.0 75 WhiB1, 50 µM WhiB1, 50 µM, alkylated 340 nm 0.090 0.08 0.06 0.04 0.02 350 400 450 50 0 55 0 600 650 0.00 300 700 350 ...

Ngày tải lên: 18/02/2014, 12:20

... 33 05 3307 Glavan F, Behm-Ansmant I, Izaurralde E & Conti E (2006) Structures of the PIN domains of SMG6 and SMG5 reveal a nuclease within the mRNA surveillance complex EMBO J 25, 51 17 51 25 Thompson ... combination of vectors: pcDNA3, pcDNA3 ⁄ IL-1+ARE, pcDNA3 ⁄ IL-1)ARE, pcDNA3 ⁄ MCPIP, MCPIP ⁄ IL-1+ARE, MCPIP ⁄ IL-1)ARE and pcDNA3 ⁄ MCPIP The pcDNA3 vector served as a control for the experiment, and ... (endogenous and exogenous) starts degradation of IL-1b mRNA and also of its own transcript For this reason, the level of mRNA for the wildtype form of MCPIP and the level of mRNA for IL-1b (Fig 5A, lane...

Ngày tải lên: 18/02/2014, 14:20

... G63 G56 D70 G57 G50 N140 R 55 R62 T131 T138 T1 45 T138 Fig (A) Superposition of UpUMPK (green) on top of E coli UMPK (blue) at the active site (B) Binding of UMP to E coli UMPK and the location of ... the backbone N of Gly63, and the phosphate group forming hydrogen bonds from O1 to Arg 55, from O2 to the backbone N and the side chain of Thr138, and from O3 to the backbone N of Gly50 A P-loop ... phosphate of 25 ⁄ The effect of orthophosphate on enzyme activity was examined It was shown that phosphate inhibited UpUMPK activity with an IC50 value of mm (Fig 7) Functional consequences of F133N and...

Ngày tải lên: 18/02/2014, 16:20

... I222 a ¼ 266 .50 b ¼ 292.92 c ¼ 307.30 0 .5 pentamers 363 7 15 124 622 19.8–3.6 0.90 (0.92) 0.13 (0.64) 4.12 (1.33) 2.9 15- fold 15. 0–3.6 1 15 706 3041 53 0 65 0.90 ⁄ 0.32 27 .5 ⁄ 32.8 0.0 15 1.68 96.7% ... is formed by an 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, ... protein of human papillomavirus 16 Mol Cell 5, 55 7 56 7 Chroboczek J, Gout E, Favier AL & Galinier R (2003) Novel partner proteins of adenovirus penton Curr Top Microbiol Immunol 272, 37 55 Galinier...

Ngày tải lên: 19/02/2014, 06:20

... and is a potent FEBS Journal 272 (20 05) 57 23 57 41 ª 20 05 The Authors Journal compilation ª 20 05 FEBS 57 35 PDGF-C and -D, structure and function L J Reigstad et al transforming growth factor of ... model of the PDGF-C growth factor domain indicates the disulfide bridges in PDGF-C to consist of Cys 250 and 294, Cys280 and 3 35, and Cys287 and 337, and the intermonomeric bonds to consist of Cys274 ... In these aggressive bone and soft tissue tumours, FEBS Journal 272 (20 05) 57 23 57 41 ª 20 05 The Authors Journal compilation ª 20 05 FEBS 57 33 PDGF-C and -D, structure and function L J Reigstad...

Ngày tải lên: 19/02/2014, 07:20

... Hamasaki, N q FEBS 2001 53 54 55 56 57 58 Characterization of calretinin domain I – II (Eur J Biochem 268) 6237 & Thorner, J (2000) Structure and calcium-binding properties of Frq1, a novel calcium ... G114, G162) [51 ,52 ] (B) GCAP-2 (G38, G72, G110, G163) [53 ,54 ] (C) Calb I –II (G29, G71) [31] (D) Nereis diversicolor sarcoplasmic Ca21-binding protein (G21, G 55, N109, G143) [55 ,56 ] (E) Calerythrin ... form of the Lipari –Szabo analysis [57 ], a global correlation time of 6.7 ^ 0 .5 ns was determined for calcium-bound 15 N-labeled CR I–II at pH 5. 4 and 310 K (Fig 7A) Also, the diffusion rate of...

Ngày tải lên: 22/02/2014, 07:20

... compilation ª 2011 FEBS L Cendron and G Zanotti CagQ (cag14/HP 053 5), CagP (cag 15/ HP 053 6) The products of these two short genes correspond to putative proteins of 126 and 114 amino acids, respectively ... a-helices and a final b-strand, F, which protrudes from the core of the monomer and runs anti-parallel to the same strand of a second monomer, allowing for the formation of the dimer The surface of interaction ... Journal compilation ª 2011 FEBS L Cendron and G Zanotti Cagd (cag3/HP 052 2) Cagd is a 55 kDa protein essential both for the secretion of CagA and the induction of IL-8 The corresponding sequence is...

Ngày tải lên: 06/03/2014, 00:21

... (pH 7 .5) to a final concentration of 10– 15 lm and filtered through a 0.22 lm Minisart filter Spectra were recorded at 25, 37 and 55 °C, and are accumulations of 16 scans recorded from 190 to 250 nm ... Journal 2 75 (2008) 58 85 58 98 ª 2008 The Authors Journal compilation ª 2008 FEBS 58 91 Glutamic acid mutants of Hsp 25 A M Morris et al Fig Chaperone activity of wild-type and mutant Hsp 25, as measured ... solution of Hsp 25 [ 75] and molecular mass An 58 94 Far-UV CD spectroscopy FEBS Journal 2 75 (2008) 58 85 58 98 ª 2008 The Authors Journal compilation ª 2008 FEBS A M Morris et al excitation wavelength of...

Ngày tải lên: 07/03/2014, 04:20