circular dichroism techniques for the analysis of intrinsically disordered proteins and domains
Báo cáo Y học: Binding of Thermomyces (Humicola) lanuginosa lipase to the mixed micelles of cis-parinaric acid/NaTDC Fluorescence resonance energy transfer and crystallographic study ppt
... the lack of a hydroxyl group of Ser146 diminishes the role of this residue in the stabilization of the substrate– ligand molecule, leading to the ÔconfusionÕ of the enzyme in the process of the ... cis-PnA/NaTDC On the other hand, these shifts might result from the quenching of the tryptophan fluorescence in the presence of the mixed micelles, which simultaneously reveals the fluorescence of the tyrosine ... Binding of TLL (and mutants) to cis-PnA and NaTDC In the presence of NaTDC, without cis-PnA The presence of NaTDC at a concentration of mM resulted in a blue shift of the kmax of the RFI of TLL...
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Báo cáo khoa học: Development of a baculovirus-based fluorescence resonance energy transfer assay for measuring protein–protein interaction potx
... in the cell culture The presence of these single fluorophore-expressing cells were essential for the FRET assay because the fluorescence of these cells served as references for the subtraction of ... single fluorophore-forming complexes are not capable of participating in FRET Instead, they increase the noise and reduce the sensitivity of FRET assays Therefore, the excess amount of these complexes ... set for the FRET assay fluorescence bleed-through and the EYFP emission resulting from the Ar-UV laser excitation) of the GFP and EYFP coexpressing cells The assay was also optimized for the measurement...
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Báo cáo sinh học: "Combined optical trapping and single-molecule fluorescence" doc
... nucleotides were placed on complementary bases, one on the 3 end of the shorter 15-mer and the other on the 5 end of the long strand; in these positions, the dye proximity causes fluorescence to be quenched ... an unzipping force of pN The dye unquenched at the point of rupture, and later bleached at t Ϸ sec See text for further details Prior to rupture, force increased monotonically, and the low light ... the bead from the center of the trap and thereby increasing the load until rupture (‘unzipping’) of the 15 base-pair duplex occurred Experiments were conducted in an assay medium consisting of...
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Tài liệu Báo cáo khoa học: DNA strand exchange activity of rice recombinase OsDmc1 monitored by fluorescence resonance energy transfer and the role of ATP hydrolysis pptx
... donor and acceptor dyes, will therefore unveil the status of physical union and separation of complementary strands during renaturation and strand exchange, respectively, and thereby assesses the ... represent the active form responsible for recombination reactions [27] The presence of ATP was found to result in the formation of helical filaments on ssDNA, whereas in the absence of ATP there was ... monitor the time course of renaturation and strand exchange activities of OsDmc1 The renaturation activity was found to be stimulated in the presence of ATP and was saturated at 1.25 lm of OsDmc1...
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Tài liệu Báo cáo khoa học: Constitutive oligomerization of human D2 dopamine receptors expressed in Spodoptera frugiperda 9 (Sf9 ) and in HEK293 cells Analysis using co-immunoprecipitation and time-resolved fluorescence resonance energy transfer pdf
... member of the D2-like family of dopamine receptors (which comprises D2, D3 and D4 receptors) These receptors are GPCRs that couple to G proteins of the Gi/o family There are two isoforms of the ... described above for whole cells and cell membranes When the effects of ligands were analysed, they were preincubated with the cells for 15 prior to the addition of the antibodies The energy transfer ... to the tag sequence These oligonucleotides were then annealed and digested with NdeI TOPOD2L and TOPOD2S were digested with EcoRI and NdeI, and the DNA fragments and the HIV tag were ligated The...
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Báo cáo khoa học: Ternary complex formation of pVHL, elongin B and elongin C visualized in living cells by a fluorescence resonance energy transfer–fluorescence lifetime imaging microscopy technique docx
... donor and acceptor, this result matches with the results from the X-ray crystallography of the VBC complex [26]; the distance between the C-terminal end of elongin B and the N-terminal end of elongin ... although the effects caused by the binding of pVHL on the 3D structure of the elongin BC complex are not exactly known The present study has clarified that conformation of pVHL and ⁄ or elongin C in the ... tumors For formation of the VBC complex, it has been elucidated that cooperation of the HSP70 and TRiC ⁄ CCT chaperone systems is required [11,12] The VBC complex further associates with cullin-2 and...
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Báo cáo khoa học: Dimerization of mammalian adenylate cyclases Functional, biochemical and ¯uorescence resonance energy transfer (FRET) studies pot
... constructs; the green box represents the GFP molecule, the 12 small black bars represent the 12 putative transmembrane segments of AC8 The names of the domains of AC8 are above the GFP/AC8 in the corresponding ... Tracking of the hydrophobic portion of AC8 (A) The left panel shows the structural diagram of 8Tm1/CFP/Tm2 The red and blue cylinders represent the transmembrane segments of the ®rst and second ... The asterisk indicates the position of a nonspeci®c antibody band The arrow heads show the positions of GFP/8Tm2C2 (c 80 kDa) and GFP/8Tm2 (c 55 kDa) The upper bands are probably oligomeric forms...
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SINGLE MOLECULE STUDY ON DNA PROTEIN INTERACTION IN PROKARYOTES AND EUKARYOTES
... tether length plus the bead radius Bead fluctuation along the y-direction is used to measure the force The extension of the tether is determined from the center of the bead to the edge of the ... temperature, 𝑘𝐵 is the Boltzmann constant, is the end-to-end extension of the tether along the force direction, and the (𝛿𝑦)2 is the variance of the transverse (perpendicular to the force direction) ... evaluated and however the location of each step is just between the steps in the fitting to be evaluated Then, the ratio between the Chi-squared of the ‘counter fitting’ and the Chi-squared of the...
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Tài liệu Báo cáo khoa học: Loose interaction between glyceraldehyde-3-phosphate dehydrogenase and phosphoglycerate kinase revealed by fluorescence resonance energy transfer–fluorescence lifetime imaging microscopy in living cells doc
... right The distances between the Nterminal amino acids of subunits O and P, subunits O and Q and subunits O and R are shown (B–D) FLIM analysis of the interaction between cerulean–GAPDH and citrine–GAPDH ... against the existence of the complex and channeling of metabolites [10,11] Thus, the existence of the complex has not been universally accepted In addition to the cytoplasmic compartment, GAPDH and ... 1.0–1.2 1.0–1.1 a The differences between the two s1 values and the two s2 values were significant (P < 0.005 for s1 and P < 0.001 for s2) differences between the two s1 values and the two s2 values...
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Báo cáo Y học: Probing intermolecular protein–protein interactions in the calcium-sensing receptor homodimer using bioluminescence resonance energy transfer (BRET) potx
... (pBRET+) by the introduction of a Ser65 fi Thr mutation in the GFP2 part of the plasmid For the construction of the c-myc-CaR and HA-CaR constructs, a MluI site was introduced after the signal ... GABAB1aEGFP and GABAB1b-EGFP parts of the original plasmids Hence, c-myc-CaR and HA-CaR consisted of the signal peptide for mGluR5, HA or c-myc and the entire CaR Construction of tagged receptors CaR and ... levels of the Rluc- and GFP2-tagged CaRs and AT1aRs are similar To evaluate the cell surface expression, we tagged HA and c-myc epitopes to the N-terminal of the CaR-GFP2 and CaR-Rluc fusion proteins, ...
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Báo cáo khoa học: Receptor- and calcium-dependent induced inositol 1,4,5-trisphosphate increases in PC12h cells as shown by fluorescence resonance energy transfer imaging pot
... activation and the reintroduction of calcium, as well as the effects of Tg on the components of these responses The peak amplitudes were calculated as the difference between signals before and after ... [20] To determine the physiological role of the calcium dependence of Ins(1,4,5)P3 metabolism, a more detailed analysis of the effects of calcium and the cross-talk between other signaling systems ... before stimulation, and were localized close to the plasma membrane As the hydrolysis of PtdIns proceeds, the subsequently formed PtdIns(4,5)P2 is converted to Ins(1,4,5)P3, and the fusion proteins...
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FUNDAMENTALS of KINEMATICS and DYNAMICS
... (bellcrank) and 7, the position of the bucket Thus, the final displacement of the bucket is controlled by two mechanisms: one is the mechanism for lifting the arm, and the other is for the rotation of the ... shape, and weight of the object all constitute input into the design process All of these will affect the size of the mechanism There are other considerations as well, such as, for example, what the ... speed of approaching point B should be The outcome of this inquiry may affect either the configuration or the type of the mechanism Within the subject of kinematics and dynamics of machines and...
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Atomistic modeling of energetics and dynamics of diffusive and frictional phenomena in c60 graphene based systems
... changing the temperature which alters the mechanism of exchanging the energy between the admolecule and the substrate We evaluate the effect of rotational degrees of freedom (DOFs) of the C60 ... of the C60/graphene system for the Hex.-In Phase configuration (b) The corresponding contour plot of the PES The potential energy profile of the path in the [100] crystallographic direction of ... coupling of adsorbate to surface 51B The substrate influences the adsorbate dynamics through the adiabatic PES of the system The difference between the value of the PES at the saddle point and at the...
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Study of pharmacokinetics of prenylflavonoids and dynamics of estrogen action in sera following ingestion of epimedium using validated, ultra sensitive cell based bioassays
... icaritin and desmethylicaritin and metabolism of these compounds, particularly, the glycosides, occurs upon the ingestion of the extract To understand the bioavailability and conversion of these ... [M−H]− and the most abundant and characteristic product ions were selected for MRM analysis The ion pairs selected for the five flavonoids and internal standard were 513→351 for icariin, 529→367 for ... respectively and internal standard in pooled rat sera, were placed at the beginning and end of each assay for determination of accuracy and imprecision (Shah et al., 2000) Accuracy and imprecision of the...
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Study of pharmacokinetics of prenylflavonoids and dynamics of estrogen action in sera following ingestion of epimedium using validated, ultra sensitive cell based bioassays 2
... on the y-axis and estradiol concentrations on the x-axis, were then fitted with the regression method that visually 90 best fits the points within the range of the test samples that maximizes the ... 175 200 [E2], pM 92 3.1.4 Isoform selectivity of DPN, PPT and genistein To assess the ER-isoform selectivity of ERα and ERβ bioassays, compounds such as, DPN, PPT and genistein, which are non-steroidal ... dihydrotestosterone and hydroxycortisone in the presence of 50 pM of estradiol for a duration of days Changes of cell culture media were carried out twice on the second and fourth day of incubation...
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Study of pharmacokinetics of prenylflavonoids and dynamics of estrogen action in sera following ingestion of epimedium using validated, ultra sensitive cell based bioassays 4
... the hydroxyl group on ring A with His-524 In contrast, for the case of icaritin, the interaction with Glu-353 and Arg-394 in the ERα ligand-binding domain was blocked due to the presence of the ... 48 h The estrogenicity of these trace amounts of unconjugated icaritin was well below the detection limits of the ERα and ERβ, as well as, MCF-7 cell-based bioassays The low amounts of the compound ... but the effects of these polymorphisms have not been widely studied The selectivity of conjugation of the flavonoids by UDP-glucuronosyltransferases depends on structure of the flavonoid and the...
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Study of pharmacokinetics of prenylflavonoids and dynamics of estrogen action in sera following ingestion of epimedium using validated, ultra sensitive cell based bioassays 5
... maintenance of bone health and other conditions requiring estrogenic action in humans, from the lab bench to bedside This thesis reported the importance and value of the development and validation of ... ex-vivo for levels of estrone and estradiol by tandem mass spectrometry, for ERα and ERβ bioactivity and MCF-7 breast cancer cell proliferative effects The concentrations of icaritin and desmethylicaritin ... initiation and progression The ability of Epimedium compounds to act as dual activators for ERα and AhR mechanistically represent a new class of SERMs These compounds should be further examined for...
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Study of pharmacokinetics of prenylflavonoids and dynamics of estrogen action in sera following ingestion of epimedium using validated, ultra sensitive cell based bioassays 6
... Sternfeld, B and Matthews, K 2006 Longitudinal analysis of the association between vasomotor symptoms and race/ethnicity across the menopausal transition: study of womens’ health across the nation ... 27008–27016 Mak, P., Cruz, F.D and Chen, S 1999 A yeast screen system for aromatase inhibitors and ligands for androgen receptor: yeast cells transformed with aromatase and androgen receptor Environ ... involved in the formation and transformation of steroid hormones in the fetal and placental compartments J Steroid Biochem Mol Biol 97(5):401-15 Peng, G., Fu, M and Zhang, D 1997 The experimental...
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Application of formal and dynamic equivalence to the translation of bị and được into english
... Bị and act as real verbs Bị and can serve as real verbs when the subject is the agent of the verb In other words, the subject of the whole sentence and the subject in the predicate are uniform ... out the application of formal and dynamic equivalence to get clear methods in translation of bị and in certain types of structures Based on the results of the survey, the types of errors and ... translation of bị and into English Therefore, this research work entails studying the functions and meanings of bị and and structures formed by bị and as well as conducting a survey for error analysis...
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