排序方式: 共有60条查询结果,搜索用时 31 毫秒
51.
O Kornysova E Machtejevas V Kudirkaite U Pyell A Maruska 《Journal of biochemical and biophysical methods》2002,50(2-3):217-232
The continuous bed technique with its attractive features, such as fritless design, one-step in situ synthesis, low back pressure and no need for pressurising the electrode vessels to suppress bubble formation was applied to form polyrotaxane-based stationary phases for capillary electrochromatography (CEC). Rotaxanes are synthesized from two classes of substances, namely linear reactive monomers and inert cyclic compounds. Upon polymerisation, a gel forms with the cyclic molecules mechanically immobilized (see Fig. 1). We have employed this simple approach, using charged derivatives of cyclodextrins in order to introduce charged groups into continuous beds and thus render them appropriate for electrochromatography. The self-assembly of supramolecular structures to form rotaxanes during the synthesis of the continuous beds is treated. The electroosmotic and chromatographic properties of the various polyrotaxane-based stationary phases synthesized are discussed, as well as the synthesis of the continuous beds, including how to affect their porosity and its influence on the efficiency of the electrokinetic separation. The applicability of the rotaxane-based continuous bed is demonstrated by separation of model compounds by reversed- and normal-phase chromatography. A separation of enantiomers is also presented. This experiment is of particular interest because it indicates that the interaction with the cavity of beta-cyclodextrin (beta-CD) is not a fundamental requirement for enantioseparations. 相似文献
52.
S Sagan O Lequin F Frank O Convert M Ayoub S Lavielle G Chassaing 《European journal of biochemistry》2001,268(10):2997-3005
Two binding sites NK-1M (major, more abundant) and NK-1m (minor) are associated with the neurokinin-1 receptor. For the first time with a bioactive peptide, the Calpha methylation constraint, shown to be a helix stabiliser in model peptides, was systematically used to probe the molecular requirements of NK-1M and NK-1m binding sites and the previously postulated bioactive helical conformation of substance P (SP). Seven Calpha methylated analogues of the undecapeptide SP (from position 5-11) have been assayed for their affinities and their potencies to stimulate second messenger production. The consequences of Calpha methylation on the structure of SP have been analysed by circular dichroism and nuclear magnetic resonance combined with restrained molecular dynamics. The decreased potencies of six out of these seven Calpha methylated SP analogues do not allow the identification of any clear-cut differences in the structural requirements between the two binding sites. Strikingly, the most active analogue, [alphaMeMet5]SP, leads to variable subnanomolar affinity and potency when interacting with the NK-1m binding site. The conformational analyses show that the structural consequences associated with Calpha methylation of SP are sequence dependent. Moreover, a single Calpha methylation is not sufficient by itself to drastically stabilize a helical structure even pre-existing in solution, except when Gly9 is substituted by an alpha-aminoisobutyric acid. Furthermore, Calpha methylation of residues 5 and 6 of SP in the middle of the postulated helix does not stabilize, but decreases (to different extents) the stability of the helical structure previously observed in the 4-8 domain of other potent SP analogues. 相似文献
53.
H Duplaa O Convert A M Sautereau J F Tocanne G Chassaing 《Biochimica et biophysica acta》1992,1107(1):12-22
Analyses of interactions between substance P (SP) and phospholipids were performed by combined surface pressure and surface potential measurements in monolayers and by 13C-NMR experiments on liposomes. This study was carried out using synthetic SP molecules: [1-13C-Gly9]SP and [1-13C-Gly2]SP. Injection of SP into the aqueous subphase led to an expansion of phosphatidylcholine (PtdCho) or phosphatidylserine (PtdSer) monolayer surface area. An apparent association constant of SP for PtdSer was estimated to be around 10(6)-10(-7) M-1. The surface potential delta V/n varied linearly with the molecular area whereas the variation of surface pressure was biphasic, suggesting that at least two binding states contributed to the monolayer expansion. These two states Si (SP is inserted into the bilayer) and Ss (SP is stuck on the surface) were observed on vesicular membranes by 13C-NMR. The kinetic of interconversion between these two states can be estimated by NMR, the Ss state being the stablest one. No perpendicular insertion of SP into these vesicular preparations seemed to occur, as previously postulated. However, SP might form aggregates in contact with these model systems, leading to a loss of permeability of the lipid vesicles. 相似文献
54.
55.
The binding of the antitumoral ellipticine derivative 2-methyl-9-hydroxyellipticinium acetate (elliptinium; NMHE) to DNA was analyzed by the combined use of DNase I footprinting and spectroscopic methods. Using two fragments of pBR322 DNA, five discrete NMHE binding sites of 5-7 protected base pairs (bp) were detected by footprinting at 4 degrees C on the analyzed regions. These corresponded to alternating pyrimidines and purines. The inactive derivative 2-methyl ellipticinium acetate L(NME) lacking a hydroxy group failed to demonstrate DNA protection even at low temperature. Ultraviolet-absorption and 1H-nmr analysis was performed using two autocomplementary octanucleotides d(TGACGTCA) (I) and d(ACTGCAGT) (II). The uv-absorption titrations resulted in an intercalative binding mode for NMHE in the oligomers. Analysis of the derived biphasic Scatchard plots yielded two binding sites corresponding to approximately 6-bp and 2-bp sizes and characterized by apparent association constants K1 approximately 10(8) M-1 and K2 approximately 10(6) M-1, respectively. The 1H-nmr analysis of exchangeable (imino) protons and nonexchangeable protons performed in the one- and two-dimensional modes confirmed the intercalation of NMHE, and further revealed the existence of multiple sites on DNA. Assuming that imino resonance line width concerned the sole kinetic effects, 10-ms order lifetimes were estimated for the drug-oligonucleotide complexes at 7 degrees C, pH 7, and 0.1 ionic strength. Finally, examination of every drug-DNA spectra in the light of the footprinting results indicated that there was a preference for binding of NMHE to the CpG (octamer I) and TpG (octamers I and II) steps. 相似文献
56.
57.
Geoffrey E. Hawkes Edward W. Randall William E. Hull Odile Convert 《Biopolymers》1980,19(10):1815-1826
The natural-abundance 15N-nuclear magnetic resonance (nmr) spectrum of the cyclic decapeptide gramicidin S has been measured and assigned in the solvents dimethyl sulfoxide, methanol, and 2,2,2-trifluoroethanol. Three methods have been investigated to distinguish between peptide groups which are exposed to or shielded from the solvent. The solvent dependence of the 15N chemical shift is correlated with the two types of peptide group in gramicidin S? those with the carbonyl group exposed or shielded from the solvent. The second method monitors the lability of the N? H proton (via the collapse of the reduced 15N-1H coupling) in the presence of added base used to promote intermolecular exchange—peptide protons shielded from the solvent exchange more slowly. The third method looks at the temperature dependence of the 15N chemical shifts in dimethyl sulfoxide. Here the data are not so distinctive as to allow the differentiation between solvent-exposed or shielded N? H bonds at all peptide groups. 相似文献
58.
Marie-Christine Maurel Odile Convert 《Origins of life and evolution of the biosphere》1990,20(1):43-48
Upon heating a dry mixture of ribose and adenine, condensation products are formed. They were identified as isomers of N6-ribosyl-adenine (Fuller, Sanchez and Orgel, 1972). Due to the current interest in nucleotide analogs as potential constituents of primitive RNA catalysts, the products were further characterized by mass spectroscopy and proton NMR. Our results fully substantiate the previous proposals. 相似文献
59.
The necessity of calcium (Ca2+) and the Ca2+-calmodulin complex for resumption and completion of meiosis, expansion of cumulus cells, viability and hyaluronidase sensitivity of in vitro cultured bovine cumulus-oocyte complexes was examined by inhibition of the Ca2+-calmodulin complex with eight graduated doses of trifluoperazine (TFP) and by Ca2+ deficiency or depletion. Doses of TFP greater than 2.5 microM decreased the percent of cumulus complexes surviving culture and oocytes completing meiosis, whereas cumulus expansion was unaffected until the cultures contained a near lethal dose (greater than 10 microM). Hyaluronidase caused dispersion of cumulus cells whenever they were expanded regardless of TFP dose. In TC-199 media the completion of meiosis I was suppressed by 0.1 to 1 mM ethylenediaminotetraacetic acid (EDTA) (P less than 0.05) and drastically reduced by 1.0 mM (P less than 0.05). Viability of the cumulus-oocyte complex was not reduced until the dose of EDTA was increased to 1.0 mM (P less than 0.0001). Cumulus expansion was also not suppressed until the dose of EDTA reached 1.0 mM (P less than 0.05). In Ca2+-free (CF) basal media Eagles, completion of meiosis I was reduced by all doses of EDTA (P less than 0.05), whereas viability of the cumulus-oocyte complex was decreased by Ca2+ deficiency or by EDTA addition to basal media Eagles (P less than 0.01). Cumulus expansion was unaffected by Ca2+ removal or chelation. In all experiments, oocytes which were not degenerate underwent germinal vesicle breakdown regardless of treatment. 相似文献
60.
Preferential conformation of substance P in solution 总被引:4,自引:0,他引:4
The three-dimensional structure of substance P has been studied by 1H-NMR, (500 MHz), and by circular dichroism (CD) in different solvents. The analysis of the different NMR parameters suggest that substance P adopts a rather extended structure in dimethylsulfoxide and pyridine. In water, besides the aggregation phenomenon, the monomeric substance P presents a complex conformational equilibrium. The addition of sodium dodecylsulfate to the aqueous solution induces, as shown by CD spectroscopy, a preferential alpha-helical conformation. And in methanol three structural conclusions may be drawn: the flexibility of the N-terminal Arg-Pro-Lys, the alpha-helical structure of Pro4-Gln5-Gln6-Phe7-Phe8 and the interaction of the C-terminal carboxamide with the primary amides from both glutamines. 相似文献