ensemble fret methods in studies of intrinsically disordered proteins

... remaining part of thsis moiety in a similar manner to the model of one of the alkyl chains in the TLL complex with di-dodecyl phosphatidylcholine [15] (not presented in Fig 6B for the sake of ... dashed lines indicate hydrogen bonds between OA and the protein (B) Comparisons between several ligand binding modes in the catalytic cleft of TLL The stereoview of OA binding modes in molecule ... micelles of cis-PnA/NaTDC The presence of mixed micelles of cis-PnA/NaTDC was also suggested by the increase in the fluorescence emission intensity of cis-PnA recorded with increasing amounts of NaTDC...

Ngày tải lên: 08/03/2014, 23:20

... for investigating the molecular mechanism involved in protein–protein interaction and for screening novel molecules that might enhance or block the protein–protein interactions of molecules of interest ... sensitive method of visualization of FRET and assessment of protein–protein interactions It can potentially allow for studying the associations of molecules of any proteins of interest Acknowledgements ... level of recombinant protein expression, manipulating it by adjusting the multiplicity of infection (MOI) By combining the merits of both FRET and the baculovirus system, a new baculovirus-based FRET...

Ngày tải lên: 30/03/2014, 20:20

... pulling 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 ... binding and stepping motion in motor proteins, or to probe molecular mechanisms underlying conformational changes We anticipate that this technique will have broad applicability to the study of ... Figure S1 A simplified diagram of the optical layout of our instrument (see also [4]) The inset is a cartoon showing a detail of the specimen in the region illuminated by the three lasers (not...

Ngày tải lên: 06/08/2014, 18:20

... function of the protein may be related to some critical steps in the conversion of inactive forms of protein to that of active forms It is interesting and intriguing to note that in the absence of ... the latter Interestingly, it appears that protein binding to ssDNA in the absence of any nucleotide cofactor results in attenuation of spontaneous annealing (compare line 1, Fig with line 2, Fig ... activity of OsDmc1 by FRET C Rajanikant et al ficant changes either in ATPase or in DNA binding domains of OsDmc1 [22] The only discernible change in OsDmc1 is an insertion of seven amino acid...

Ngày tải lên: 19/02/2014, 07:20

... changes in the proteins rather than changes in protein– protein interaction A combination of several methods seems therefore important for the demonstration of GPCR oligomerization The D2 dopamine ... of signal Membranes containing the equivalent of 100 fmol of receptors (as labelled with [3H]spiperone) were incubated with 2.5 nM Eu3+-labelled anti-FLAG Ig, in a total volume of 500 lL of incubation ... binding of ligands to the D2 dopamine receptor may exhibit co-operativity, which can be accounted for in terms of interactions between binding sites in an oligomer There is also the question of...

Ngày tải lên: 21/02/2014, 00:20

... change of elongin C induced by binding of elongin B Increased FRET signals between pVHL-cerulean and citrine-elongin C by coexpression of elongin B suggest that a conformation change of elongin C induced ... fluorescent proteins in the following four combinations: cerulean-elongin B and citrineelongin C; cerulean-elongin B and elongin C-citrine; elongin B-cerulean and elongin C-citrine; and elongin B-cerulean ... lifetimes of donor proteins using time-domain FLIM Using the FRET FLIM technique, we observed strong intermolecular FRET signals between elongin B and elongin C For stable binding of pVHL30 to elongin...

Ngày tải lên: 16/03/2014, 05:20

... calmodulin, binding to the C-terminus [22] Issues of speci®city of intermolecular interactions were addressed with AC5 or AC6, which are inhibitable by Ca2+, apparently independently of calmodulin ... to play a dominant role in the intermolecular interaction The more discerning technique of FRET analysis in live cells showed that in addition to interacting with Tm1 and traf®cking to the plasma ... Rescue of inactive mutants by half molecules of AC8 in vivo A corollary of the experiments described above involving dominant negative suppression of adenylate cyclase activity is the rescue of inactive,...

Ngày tải lên: 23/03/2014, 21:21

... without forming aggregates Many kinds of proteins are involved into the DNA compaction process Histone chaperons are a group of proteins in eukaryotic cells to help chromatin assembly The binding mechanisms ... concentration of the reaction buffer into infinite or to add certain amount of competing DNA, both of which are designed to obtain the optimal ratio of DNA to histone proteins As described in the previous ... regained great interests due to their functions in facilitating reconstitution of chromatin for functional and structural studies in vitro Histone chaperones are a group of proteins which can bind...

Ngày tải lên: 12/10/2015, 17:34

... fluorophores, being independent of the strength of protein–protein interactions In the case of very weak interactions as described in this article, rapid association and dissociation of FRET pairs may ... depend on the length of linkers In the linker containing seven amino acids, a Pro-Pro sequence that is not contained in other linkers is incorporated This less flexible structure may inhibit fluorophores ... because of its inherent instability Our FRET FLIM system may serve as a valuable tool for investigating weak interactions in the complex in living cells C-3¢, and pcerulean-C1 For the construction of...

Ngày tải lên: 16/02/2014, 09:20

... demonstrate agonist-induced alterations in BRET signal intensities reflecting 7TM dimer rearrangement as a result of the activating twist in the ATDs of the CaR homodimer Further investigations into the ... forms used in other studies [34,40] In the window of 500–530 nm the emission of Rluc/GFP2 transfected cells was  7.4 times higher than that of Rluc transfected cells (Fig 6A) Interestingly, the ... this BRET2 assay in contrast to the original BRET assay using Rluc/coelenterazine h [34,40], may hold some practical advantages for future studies of GFP fusion proteins As the obtained BRET signal...

Ngày tải lên: 08/03/2014, 09:20

... following an increase in Ins(1,4,5)P3 This has been observed in Madin–Darby canine kidney (MDCK) cells, in which Ins(1,4,5)P3 injection caused the redistribution of fusion proteins, regardless of ... containing CCh, which induced a second calcium increase and a further increase in Ins(1,4,5)P3 (Fig 3Aa) Both of these calcium increases were accompanied by increased Ins(1,4,5)P3, indicating ... fusion proteins changes almost instantaneously following the disruption of PtdIns(4,5)P2 binding Therefore, the delay observed in our previous study indicates that the release of fusion proteins...

Ngày tải lên: 30/03/2014, 03:20

... type of constraining condition determines the number of degrees of freedom (DOF) a mechanism has If the constraining condition allows only one DOF between the two links, the corresponding joint ... tipping of the dump bed All the previous examples involved only links with two connections to other links Such links are called binary links In the example of Figure 1.6, in addition to binary links, ... its number of degrees of freedom This translates into a number of independent input motions leading to a single follower motion A single unconstrained link (Figure 1.7a) has three DOF in planar...

Ngày tải lên: 08/04/2014, 11:35

Ngày tải lên: 06/10/2014, 15:21

... dependence of kinetic nanofriction We use the Einstein’s theory of Brownian motion to analyze the surface diffusion of the C60 admolecule on graphene, and show that the decrease of kinetic nanofriction ... regimes of surface diffusion in C60/graphene system according to the effect of temperature and rotational degrees of freedom (DOFs) of the admolecule In the case of rotational C60 (the set of arrows ... determine the kinetic nanofriction of mobile adsorbates [72] The Einstein’s theory of Brownian motion, which is widely applied in microsystems, provides a link between the diffusion coefficient of...

Ngày tải lên: 08/09/2015, 17:49

... using lipofectamine Sixteen hours post-transfection, the cells were washed, trypsinized and split at ratio of in 10, in 100 and in 1000 into selective media containing 100 μg/mL of hygromycin ... contained 143.3 mg/g of icariin, 0.157 mg/g of icariside I, 18.68 mg/g of icariside II, 0.009 mg/g of icaritin, and 0.122 mg/g of desmethylicaritin, respectively Following Epimedium administration, ... human subject in this study consumed 500 mL of decoction containing icariin: 12.6 mg, icaritin: 0.75 mg, and desmethylicaritin: 0.19 mg A single dose of 50 g per subject was chosen according to calculations...

Ngày tải lên: 10/09/2015, 15:48

... Combinatorial effects of binary mixtures of estradiol with genistein and Epimedium prenylflavonoids To study combinatorial effects of flavonoids and estradiol, binary mixtures containing increasing ... cell line 101 Figure 17: ERα combinatorial effects of binary mixtures of estradiol with genistein and Epimedium prenylflavonoids ERα stable cells were incubated with increasing doses of genistein ... 18: ERβ combinatorial effects of binary mixtures of estradiol with genistein and Epimedium prenylflavonoids ERβ stable cells were incubated with increasing doses of genistein (A), icariin (B), icariside...

Ngày tải lên: 10/09/2015, 15:54

... derivatives, genistin and naringin not bind to the sex 201 hormone-binding globulin Hence, it is not likely that icaritin will bind to the sex hormone-binding globulin protein found in serum The late ... absence of serum, binding of quercetin to recombinant ERα was comparable to that of genistein and kaempferol (Maggiolini et al., 2004; Leung et al., 2004) The presence of serum reduced ERα binding of ... human serum albumin but not sex hormone-binding globulin, which would influence its bioavailability A large proportion of proteins found in serum are made up of albumin, which binds and transports...

Ngày tải lên: 10/09/2015, 15:54

... ascertain whether the ingestion of estrogenic prenylflavonoids from Epimedium for the purpose of maintaining bone health can increase risk of ERα-positive breast cancer Interestingly, icaritin, ... (-)-epigallocatechin-3-gallate significantly increased the biochanin A area under the plasma concentration versus time curve in rats in both intravenous and oral administration of biochanin A According to ... especially in terms of icariin but it had a lesser amount of icaritin - 143.3 mg/g icariin, 0.009 mg/g icaritin and 0.122 mg/g desmethylicaritin The amounts of icariside I and icarside II were determined...

Ngày tải lên: 10/09/2015, 15:54

... Ma, D.F., Qin, L.Q., Wang, P.Y and Katoh, R 2008 Soy isoflavone intake increases bone mineral density in the spine of menopausal women: meta-analysis of randomized controlled trials Clin Nutr 27(1):57–64 ... Pharmacokinetics and bioavailability of the bioflavonoid biochanin A: effects of quercetin and EGCG on biochanin A disposition in rats Mol Pharm 4(6):865-72 Mueller, S.O 2002 Overview of in vitro ... determination of serum estrogenic bioactivity in children J Clin Endocrinol Metab 87:791–797 Pasqualini, J.R 2005 Enzymes involved in the formation and transformation of steroid hormones in the...

Ngày tải lên: 10/09/2015, 15:54