immobilization of proteins for single molecule fluorescence resonance energy transfer measurements of conformation and dynam
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
Ngày tải lên :
08/03/2014, 23:20
... bonds of D92 and H110 with carboxyl group of the OA molecule, specific for this particular ligand conformation Whether this form of OA binding is enhanced further by the more stable conformation of ... composed of all the atoms of all the residues between E1 and L269 in the case of molecule A (chain A) and molecule ˚ B (chain B), with a rmsd of 0.17 A between the corresponding Ca atoms of molecule ... density in the molecule B in comparison with higher disorder of the ligand in the molecule A However, despite of the lack of strong lattice contacts that may affect the conformation of the lid,...
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Báo cáo khoa học: Development of a baculovirus-based fluorescence resonance energy transfer assay for measuring protein–protein interaction potx
Ngày tải lên :
30/03/2014, 20:20
... 201–207 24 Selvin, P.R (1995) Fluorescence resonance energy transfer Methods Enzymol 246, 300–334 25 Wu, P & Brand, L (1994) Resonance energy transfer: methods and applications Anal Biochem 218, ... allow for studying the associations of molecules of any proteins of interest Acknowledgements We acknowledge Dr Daryl Webb for his excellent technical assistance with confocal microscopy; Professor ... coexcitation of EYFP, and a minimum energy level was used for imaging to minimize photobleaching Optimum emission windows for both GFP and EYFP were determined with the spectral scanning program of a...
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Báo cáo sinh học: "Combined optical trapping and single-molecule fluorescence" doc
Ngày tải lên :
06/08/2014, 18:20
... acids and polypeptides, or conformational rearrangements associated with the translocation of processive enzymes Moreover, simultaneous optical trapping and single- molecule fluorescence measurements ... simultaneous singlemolecule fluorescence and optical trapping The choices of dyes and excitation wavelengths were optimized to minimize two-photon excitation and achieve acceptably long fluorescence ... wave (singlesided total internal reflection fluorescence, TIRF) Prior attempts to marry optical trapping with single- molecule fluorescence failed to achieve spatially coincident fluorescence and...
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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
Ngày tải lên :
19/02/2014, 07:20
... between 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 ... labeled Phi-W and rhodamine labeled Phi-C (1) with 1.25 lM of OsDmc1 in absence of ATP; (2) with 1.25 lM of OsDmc1 in presence of 2.0 mM of AMP-PNP; (3) with 1.25 lM of OsDmc1 in presence of 2.0 mM ... further, and the strand exchange property of OsDmc1 was monitored by pairing an unlabeled Phi-C single strand with duplex oligonucleotide formed by complementary annealing of FRET dye labeled strands...
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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
Ngày tải lên :
21/02/2014, 00:20
... bands (at 39 and 80 kDa) for D2S, but only one band (85 kDa) for D2L receptors (Fig 3, lane 3) The smaller band (39 kDa) observed for D2S may correspond to a disruption of an oligomeric form of ... R (2002) Monitoring of ligand-independent dimerization and ligand-induced conformational changes of melatonin receptors in living cells by bioluminescence resonance energy transfer J Biol Chem ... means of secondary conjugated antibodies Anti-gp120 Ig and anti-FLAG Ig identified bands corresponding to proteins with a molecular mass equivalent to 43 kDa and 85 kDa for D2L and 39 kDa and...
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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
Ngày tải lên :
16/03/2014, 05:20
... unambiguous evidence for spatial and temporal interactions between proteins and conformational changes of proteins occurring in living cells The occurrence of FRET can be accurately and finely determined ... pVHL-cerulean and citrine-elongin C by coexpression of elongin B suggest that a conformation change of elongin C induced by binding of elongin B may occur and that this conformational change of elongin ... clarified that conformation of pVHL and ⁄ or elongin C in the absence of elongin B was different from that in the VBC complex and that conformation of elongin C was changed upon binding of elongin...
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Báo cáo khoa học: Dimerization of mammalian adenylate cyclases Functional, biochemical and ¯uorescence resonance energy transfer (FRET) studies pot
Ngày tải lên :
23/03/2014, 21:21
... cells was measured for after adding, 20 lM forskolin and 10 lM prostaglandin E1 for transfections involving AC5 and AC6; 20 lM forskolin, 10 lM prostaglandin E1 and mM Ca2+ for transfections involving ... with the formation of homomultimeric complexes of AC8 molecules We wondered whether a similar approach might reveal that heteromultimeric complexes could form between different isoforms of adenylate ... nM for min) to activate CCE cAMP accumulation in the intact cells was measured for after adding, vehicle (Basal); 20 lM forskolin (Forsk); 20 lM forskolin and mM Ca2+ (Forsk/ Ca2+); or 20 lM forskolin,...
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SINGLE MOLECULE STUDY ON DNA PROTEIN INTERACTION IN PROKARYOTES AND EUKARYOTES
Ngày tải lên :
12/10/2015, 17:34
... Professor YAN Jie (PhD), for providing me the opportunity to work in Biophysics and Single Molecule Manipulation Lab and his professional guidance, mentorship and numerous help in my research and ... 59 iii Abstract Single molecule manipulation techniques have become versatile tools to study the force and motions of biological molecules such as DNA and proteins, and DNA-protein interaction ... resolution of nanometers and force resolution of piconewtons To date, the widely-used single molecule tools include optical tweezers (OT), atomic force microscopes (AFMs), and magnetic tweezers (MT), of...
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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
Ngày tải lên :
16/02/2014, 09:20
... Determination of fluorescence resonance energy transfer (FRET) between two fluorescent proteins using fluorescence lifetime imaging microscopy (FLIM) is a technique for the observation of protein–protein ... in the presence of citrine–GAPDH by energy transfer (Fig 2B) Short and long lifetimes, s1 and s2, were reduced to 1.06 and 3.13 ns, respectively In order to examine the stability of the instrument, ... expressing cerulean–GAPDH and citrine–GAPDH after sequential measurements of donor and acceptor fluorescence (Fig 2C) Perfectly overlapped decay curves for the first and second measurements FEBS Journal...
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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
Ngày tải lên :
08/03/2014, 09:20
... fluorescence resonance energy transfer and bioluminescence resonance energy transfer J Biol Chem 276, 14092–14099 Xu, Y., Piston, D.W & Johnson, C.H (1999) A bioluminescence resonance energy transfer ... One explanation for the lack of agonist-induced BRET for CaR is that the chromophore/fluorophore of the Rluc and GFP molecules are positioned so close in the resting conformation of the homodimer ... characterization of wild type (WT) AT1a receptor and AT1aD359-GFP2 was performed analogously, except that HEK 293 cells were used instead of tsA cells Fluorescence and luminescence measurements For the measurements...
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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
Ngày tải lên :
30/03/2014, 03:20
... the products of PtdIns metabolism and change their intracellular localization Recently, a fluorescence resonance energy transfer (FRET)-based assay of the redistribution of PHD fusion proteins from ... the PHD fusion proteins of the cyan and yellow variants of GFP (cyan fluoresent protein CFP–PHD and yellow fluoresent protein YFP–PHD) bound to PtdIns(4,5)P2 before stimulation, and were localized ... removal of calcium entry, we compared the averaged time-dependent restoration plots for Ins(1,4,5)P3 and calcium following removal of extracellular calcium and removal of both calcium and carbachol...
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FUNDAMENTALS of KINEMATICS and DYNAMICS
Ngày tải lên :
08/04/2014, 11:35
... subject of kinematics and dynamics of machines and mechanisms can be taught I have taught kinematics and dynamics of machines and mechanisms for many years and have always felt that concepts and ... FUNDAMENTALS of KINEMATICS and DYNAMICS of MACHINES and MECHANISMS 0257/FM/Frame Page Friday, June 2, 2000 6:38 PM FUNDAMENTALS of KINEMATICS and DYNAMICS of MACHINES and MECHANISMS Oleg ... kinematics and dynamics of the offset slider-crank mechanism, with plots and animation • fourBar.nb provides a complete solution of the kinematics and dynamics of the four-bar mechanism, with plots and...
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Atomistic modeling of energetics and dynamics of diffusive and frictional phenomena in c60 graphene based systems
Ngày tải lên :
08/09/2015, 17:49
... crucial for developing high performance NEMS, and for guiding self-organized patterns and structures Recently, a graphene Moiré pattern was employed to trap and construct arrays of C60 molecules for ... Potential energy diagram for the adsorbate moving laterally (parallel to the surface) The activation energy of diffusion Ea, is equal to the energy difference of the minima of curves and in (b) ... required for better understanding of the motion of a variety of molecules on graphene-based substrates, which will be used in future applications 1.3.2 Physisorption of fullerenes on graphene and...
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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
Ngày tải lên :
10/09/2015, 15:48
... nM of icariin, icaritin and desmethylicaritin; 5, 30 and 60 nM of icariside I and 71 icariside II, respectively and internal standard in pooled rat sera, were placed at the beginning and end of ... with 45% of solvent B for before sample injection The gradient was programmed to increase from 45% to 90% of solvent B in and maintained at 90% of B for to complete the separation Standards or ... estimation of the LOD and LOQ involved analyzing decreasing concentrations of analytes The LOD was defined as having a S/N of 3:1, and a S/N of 10:1 was defined for LOQ (U.S Food and Drug Administration,...
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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
Ngày tải lên :
10/09/2015, 15:54
... 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 ... 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 Similar to what was observed for ERα and ERβ ... (0.0158±0.0057) and y= (1.9082±0.0836)x + (0.0183±0.0068), with R2 of > 0.994 and > 0.998 for icaritin and desmethylicaritin respectively The LOD and LOQ for icaritin were estimated to be 11 pM and 33...
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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
Ngày tải lên :
10/09/2015, 15:54
... determined to be 8.1% and 16% respectively Detection limits for estradiol in ERα and ERβ bioassays were 8.4 and 13.1 pM, respectively This set of assays is simple and rapid to perform and offers highthroughput, ... separately of estradiol valerate and Epimedium pubescens decoction Serum samples were obtained and assayed ex-vivo for levels of estrone and estradiol by tandem mass spectrometry, for ERα and ERβ ... differentiation of ERα and ERβ bioactivities We validated this aspect of our bioassays using well-known ER-isoform selective ligands, such as genistein, PPT and DPN Similar to the results reported by Escande...
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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
Ngày tải lên :
10/09/2015, 15:54
... obtained and assayed 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 ... MCF-7 breast cancer cells of pure compounds and for measurement of global estrogenicity of complex mixtures To enable the study of the whole spectrum of SERM activities of Epimedium compounds on ... duration of two studies and the number of metabolites that were examined also differed The rat study reported the levels of total amounts of prenylflavonoids absorbed (sum of conjugated and unconjugated...
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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
Ngày tải lên :
10/09/2015, 15:54
... 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 ... McDaniel, T.K and Chee, M.S 2004 A novel, high-performance random array platform for quantitative gene expression profiling Genome Res 14(11):2347-56 234 Kuiper, G.G., Carlsson, B., Grandien, K., ... 2005 Pharmacology of estrogens and progestogens: influence of different routes of administration Climacteric Suppl 1:3-63 Kuo, M.L and Yang, N.C 1995 Reversion of v-H-ras-transformed NIH 3T3 cells...
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