selection of human vh single domains with improved biophysical properties by phage display
Tài liệu Báo cáo khoa học: Selection of stably folded proteins by phage-display with proteolysis docx
... 2004 Selection of folded proteins by phage- display (Eur J Biochem 271) 1611 Fig Effect of structural change on the selection (A) Structure of cyt b562 Residues M7 and H102 are the ligands of heme ... result of selection If the cutting sites are in the flexible loop of the native structure, they could prevent the selection of stable proteins By examining the structures of the proteins studied by ... was substituted with fragmented genomic Escherichia coli DNA The phage library was then challenged by several proteases Table Summary of proteins studied using phage- display with proteolysis...
- 6
- 444
- 0
Báo cáo khoa học: Cloning, expression and interaction of human T-cell receptors with the bacterial superantigen SSA ppt
... subsets are expanded by interaction with SSA Some authors indicated clonal expansion of T-cells bearing human Vb3, Vb12, Vb17 and Vb19 [20] Others showed proliferation of human T-cells bearing ... also analyzed by immunoblotting using an anti-His mAb (Sigma) or a rabbit anti-SSA Rabbit polyclonal antisera were obtained by immunization with mgÆmL)1 of SSAia mixed with a volume of complete ... SSA by ELISA with negative results, and diluted with RPMI 1640 or NaCl/Pi (1 : 1, v/v) Blood samples were taken with the understanding and written consent of each subject Twenty millilitres of...
- 9
- 485
- 0
Báo cáo khoa học: Reaction of human UMP-CMP kinase with natural and analog substrates docx
... mg protein for mL of gel), washed with 50 mL of buer A and 10 mL of buer A containing 0.3 M NaCl, before elution of the protein with buer A containing M NaCl Further purication by gel ltration ... elution volume of UMP-CMP kinase indicated a Stokes radius of 2.8 nm, in agreement with a monomer species Expression and purication of human UMP-CMP kinase as a fusion protein with GST BL21 gold ... groups with catalytic efciencies in the range of 107 M)1ặs)1 for UMP and CMP, and of 105 M)1ặs)1 for AraCMP, L-3TCMP and dCMP (Table 1) Careful characterization of the reaction catalysed by human...
- 7
- 384
- 0
Báo cáo khoa học: Functional association of human Ki-1⁄57 with pre-mRNA splicing events docx
... effect of EGFP–Ki-1 ⁄ 57 in modifying the pattern of splicing of E1A mRNA in comparison with empty pEGFP vector (Fig 4B) Expression of EGFP– Ki-1 ⁄ 57 leads to formation of the 10S and 9S isoforms, ... isoforms: 13S, 12S, 11S, 10S, and 9S (Fig 4A) [22] These isoforms can be monitored by RT-PCR followed by agarose gel analysis, where the intensity of each band in the gel directly correlates with ... seemed to be isoform specific for each Ki-1 ⁄ 57 region, as the formation of 10S mRNA was only increased by the C-terminal region of Ki-1 ⁄ 57 although with a lower efficiency in comparison with the...
- 14
- 369
- 0
Báo cáo y học: "Rev and Rex proteins of human complex retroviruses function with the MMTV Rem-responsive element" doc
... Activity of HIV-1 Rev on the MMTV RmRE in human cells A Reporter activity of RevGFP on the RmRE in Jurkat T cells Cells were electroporated with pHMRluc or pHMΔeLTRluc (1 μg) with either 20 μg of EGFP, ... function in human cells The reports of MMTV infection of human cells and detection of MMTV sequences in breast cancers and lymphomas [14-18] appear to be feasible since most steps of viral replication ... enhancement of luciferase activity by Rem, Rev and Rex in human T cells No effect of Rem was observed on the HIV and HTLV response elements (Figure 8) Surprisingly, the Rec export protein from the human...
- 13
- 246
- 0
Characterization of the interaction of EEN and its domains with ca2+ and proline rich domain
... Spectrum of BAR domain of EEN 33 Figure 4.2.1 B: SDS PAGE of BAR domain of EEN after FPLC 33 purification v Figure 4.2.2 Multi TOF MS of BAR domain 34 Figure 4.2.3 CD spectrum of BAR domain of EEN ... of recombinant protein 16 3.1.1 Vector Design 16 3.1.2 Cloning of EEN full length and its domains 18 3.1.3 SH3p11 cloning system 19 3.2 Expression of EEN and its domains 20 3.3 Purification of ... structure prediction of BAR domain 35 of EEN using SWISS-MODEL Figure 4.3.1 FPLC UV spectrum of the random coil domain of 37 EEN Figure 4.3.2 FPLC UV spectrum of ΔBAR domain of EEN 37 during purification...
- 90
- 280
- 0
Disulfide Bond Formation of Heterodimer and Heterotrimer of Human Laminin332 Coiledcoil Domains
... normally Fig Failure of disulfide bond formation by a single mutation of cysteines C1 and C2 at the N-terminus of LCC domains Two normal LCC domains of α3, β3, and γ2 with one of the single cysteine ... Results 3.1 Mutation of cysteine residues of LCC domains of human LM-332 into alanine residues by site-directed mutagenesis One cysteine residue at the C-terminus of β and γ LCC domains and two cysteine ... N-terminus of LCC domains with mutations of cysteines C1 and C2 We co-expressed one single cysteine mutant among α3C1, α3C2, β3C1, β3C2, γ2C1, or γ2C2 with two normal LCC domains of α3, β3, or...
- 8
- 227
- 0
Báo cáo khoa học: Effects of cardiomyopathic mutations on the biochemical and biophysical properties of the human a-tropomyosin docx
... transition, either because of lack of resolution or because the transition was invisible to this probe Thus, the analysis of the melting profile of human Tm was enhanced by the use of different probes ... of the actomyosin S1 Mg2+ ATPase activity by Tm in the presence of troponin The results are expressed as a percentage of the actin-activated Mg2+ ATPase of myosin S1 obtained in the absence of ... increased, however, they WT Ala63Val Biophysical properties of mutant tropomyosins The effect of the mutations on the overall stability of the proteins was evaluated by circular dichroism (CD) and...
- 9
- 603
- 0
Báo cáo khoa học: Inhibition of human MDA-MB-231 breast cancer cell invasion by matrix metalloproteinase 3 involves degradation of plasminogen docx
... c.p.m.) were mixed with samples in a buffer containing 20 mM Tris, 200 mM NaCl and mM CaCl2 (pH 8.0) in a final volume of 200 lL with or without the MMP activator APMA (1 mM) or with or without EDTA ... been previously degraded by MMP-3, then activated by t-PA (100 ng) in the presence of type IV collagen cofactor, or by partially purified MDAMB-231 cell membranes containing single- chain urokinase ... substrate-degrading capacity of intact plasminogen (3 lg) activated by t-PA (100 ng) in the presence of collagen type IV cofactor, or by cell membrane-associated PAs (equivalent of 100 ng scuPA) but...
- 8
- 320
- 0
Báo cáo Y học: Substrate specificity of human kallikrein 2 (hK2) as determined by phage display technology docx
... selected by hK2 using phage display substrates thus confirming the success of phage display substrate selection Finally, a SwissProt database search with selected sequences identified three potential human ... library with hK2 showed that no phage was in duplicate which is in contrast to selections with other types of phage display libraries (antibody fragments, ligands, or peptide binders) where selections ... reported the extent of diversity of the library used in the screening, this being a direct product of the number of different combinations of amino acids displayed by the phage Cloning substrates...
- 8
- 411
- 0
Báo cáo hóa học: "Synthesis of YVO4:Eu3+/YBO3 Heteronanostructures with Enhanced Photoluminescence Properties" pptx
... were collected by centrifugation Preparation of Sol–Gel Solution To 100 mL of water–ethanol solution (the volume ratio is 1:4) 3.83 g of Y(NO3)3Á6H2O and 0.68 g of H3BO3 (*10 mol% of excess) were ... exhibit two series of XRD patterns, namely, those of YVO4 and YBO3 In addition, the intensities of the peaks of YBO3 increase with the shell ratio Figure shows enlarged XRD patterns of some typical ... nanocrystals by the sol–gel process Figure 3c is HRTEM image of a single particle of the heteronanostructures (SR = 1/2) Interestingly, two lattice fringes of different spacing appear in a single nanoparticle...
- 6
- 312
- 0
Atomic force microscope imaging of human red blood cells and theri mechanical properties
... schematic illustration of the method of operation of an AFM The position of the cantilever of the AFM is controlled by three piezo-ceramic controllers, which place the tip of the AFM in the x and ... 3D view of a fresh RBC 48 Figure 22 Cross section profile of a dehydrated RBC 50 Figure 23 Force curves of A, B, C, D, and E of the dehydrated RBC 51-53 Figure 24 Young’s modulus of points ... modulus of points A, B, C, D, and E of a swollen RBC in PBS solution 59 Figure 28 Cross section profile of a fresh RBC in PBS solution 60 Figure 29 Force curves of A, B, C, D, and E of a fresh...
- 88
- 117
- 0
Báo cáo khoa học: Identification of calreticulin as a ligand of GABARAP by phage display screening of a peptide library pdf
... displacement of CRT from GABARAP by the binding of N1 to GABARAP, suggesting the direct competition of the peptide with CRT for a common binding site on natively folded GABARAP Discussion Phage display of ... ligand The phage display screening did not result in a single dominating peptide sequence The considerable sequence diversity of our phage display selected peptides was used to our advantage by acknowledging ... sequence alignment of phage display selected peptides with highest similarity to N1 Phage display selected peptide N1 binds to GABARAP We quantitatively investigated the binding of fluorescein-labelled...
- 13
- 560
- 0
Studies on the antibody repertoire in a dengue virus immune subject and isolation of neutralizing antibodies by phage display technology
... 4°C with addition of 0.02% Sodium Azide, or at -80°C with addition of 15% Glycerol For input tittering, 50 µL of XL-1 Blue culture was infected with 1µL of 10-7, 10-8 or 10-9 dilution of the phage ... The versatility of phage library design to display peptides or proteins of varying length and complexity (42, 43) The key steps in the selection of specific antibodies from phage display libraries ... homogenate of 65 infected Ae.aegypti, detected by Immunofluorescence Figure 19 Generation of hybridoma and construction of the chimeric 68 human/ mouse EDIII phage library Figure 20 Binding of 3H5...
- 122
- 342
- 0
Create pictures worth framing with this selection of tips 3
... background of thinking everything has to be even and centered Though the pursuit of perfection maybe a noble one, it is not needed in the realm of photography Capture a more interesting picture by having ... goals as you pursue your passion By following the foregoing tips, you can learn to make the most of your natural talent, and turn a slew of amateur photos into a professional portfolio http://www.wedding-photography-tampa.com/ ... picture by having your subject be off to the side of the frame If your camera has an auto-focus feature, it may try to lock onto whatever appears in the middle of the frame Focus manually and lock...
- 3
- 238
- 0
Tài liệu Báo cáo khoa học:Tyrosine phosphorylation of tau regulates its interactions with Fyn SH2 domains, but not SH3 domains, altering the cellular localization of tau ppt
... interaction of tau with Fyn-SH3, in line with our previous work [21] These findings, however, contrast with those of a previous study in which a deletion mutant of tau lacking residues 169–179 displayed ... cotransfected with plasmids expressing V5-tagged human 2N4R tau, the longest isoform of tau present in the adult human CNS, and Fyn Following transfection, CHO cells were treated either with pervanadate, ... binding Tyr18 of tau plays an important role in the interaction of tau with Fyn-SH2 Fig Tyrosine phosphorylation of tau increases its association with Fyn-SH2 CHO cells were cotransfected with plasmids...
- 11
- 628
- 0
Tài liệu Báo cáo khoa học: The crystal structure of human a-amino-b-carboxymuconatee-semialdehyde decarboxylase in complex with 1,3-dihydroxyacetonephosphate suggests a regulatory link between NAD synthesis and glycolysis ppt
... design of enzyme inhibitors with potential medical interest Crystal structure of human ACMSD C-ter N-ter Results and Discussion Overall quality of the model The three-dimensional structure of hACMSD ... orientation of Arg47, and is ‘edge-on’ oriented with respect to the DHAP aliphatic ˚ chain with a shortest distance of 3.7 A from C1 (Fig 3B) Such a conformation is compatible with establishment of an ... with ˚ Arg47 (distance of 3.0 A), an electrostatic interaction 2+ ˚ ion (closest distance of 4.1 A) and an with the Zn extensive network of hydrogen bonds involving a set of well-ordered solvent...
- 9
- 796
- 0
Tài liệu Báo cáo khoa học: Mapping the functional domains of human transcobalamin using monoclonal antibodies pptx
... with the maximal mAb heparin effect on the functional activity of TC Arrows show the hypothetical movement of the domains after attachment of Cbl, see the main text of the receptor related domains ... TC in 1.8 mL of human serum was labeled with the radioactive ligand (300 pm of 57Cbl, 30 min) For the experimental sample, 850 lL of the radiolabeled serum was mixed with 20 lL of TC fragment ... pattern of immunoreactive bands by western blotting of TCp was similar within the group mAbs (3-9, Q2-2 and TC4, see the corresponding lanes in Fig 4B) These mAbs reacted with the whole set of TCp...
- 12
- 514
- 0
Tài liệu Báo cáo khoa học: Insights into the interaction of human arginase II with substrate and manganese ions by site-directed mutagenesis and kinetic studies Alteration of substrate specificity by replacement of Asn149 with Asp docx
... reactions were followed by measuring the formation of ornithine, determined by the method of Chinard [42] Protein concentrations were determined by the method of Bradford [43], with BSA as standard ... of the arginase–manganese interaction was significantly altered by replacement of His120 with asparagine This aspect was quantitatively evaluated by following the Mn2+-dependent reactivation of ... analysis of the effect was not performed, the inhibition by 20 mm agmatine was eliminated by saturation with l-arginine, indicating the competitive character of the inhibition produced by the substrate...
- 9
- 651
- 0
Báo cáo khoa học: Binding of ATP at the active site of human pancreatic glucokinase – nucleotide-induced conformational changes with possible implications for its kinetic cooperativity doc
... concentration of 0.5 lgÆlL)1 and a GST–hGK ⁄ trypsin ratio of 400 : (by mass) The effect of preincubation with the substrates Glc (40 mm) or MgATP (2 mm ATP ⁄ mm MgAc) on the extent of proteolysis of WT ... ATP binding at active site of human glucokinase J Molnes et al human and yeast hexokinases, the hydroxyl group of this conserved Thr interacts with the a-phosphate of ATP [21,23,24], and a Thr ... determined by its enhancement of the ITF signal (Table 2) In the absence of AdNs, a hyperbolic binding isotherm for Glc was observed, with a Kd value of 4.2 ± 0.1 mm at 25 °C Titration with Glc...
- 15
- 374
- 0
Tìm thêm: hệ việt nam nhật bản và sức hấp dẫn của tiếng nhật tại việt nam xác định các mục tiêu của chương trình khảo sát chương trình đào tạo của các đơn vị đào tạo tại nhật bản khảo sát chương trình đào tạo gắn với các giáo trình cụ thể xác định thời lượng học về mặt lí thuyết và thực tế điều tra đối với đối tượng giảng viên và đối tượng quản lí điều tra với đối tượng sinh viên học tiếng nhật không chuyên ngữ1 khảo sát các chương trình đào tạo theo những bộ giáo trình tiêu biểu nội dung cụ thể cho từng kĩ năng ở từng cấp độ phát huy những thành tựu công nghệ mới nhất được áp dụng vào công tác dạy và học ngoại ngữ mở máy động cơ lồng sóc mở máy động cơ rôto dây quấn các đặc tính của động cơ điện không đồng bộ hệ số công suất cosp fi p2 đặc tuyến hiệu suất h fi p2 đặc tuyến mômen quay m fi p2 đặc tuyến tốc độ rôto n fi p2 động cơ điện không đồng bộ một pha sự cần thiết phải đầu tư xây dựng nhà máy từ bảng 3 1 ta thấy ngoài hai thành phần chủ yếu và chiếm tỷ lệ cao nhất là tinh bột và cacbonhydrat trong hạt gạo tẻ còn chứa đường cellulose hemicellulose