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1.
The conformational transition and the cation-binding properties of aqueous furcellaran (a gel-forming, low-sulfated polysaccharide of the carrageenan family) in various salts and salt mixtures was studied by optical rotation and by 133Cs-nmr. The results were compared with theoretical predictions based on the Poisson–Boltzmann cell model (PBCM). The conformational transition of furcellaran occurs in a single step, which implies a nonblocklike distribution of sulfate groups along the polymer chain. The chloride salts of sodium, lithium, and tetramethylammonium are equally potent in inducing helix formation of furcellaran, indicating that these ions act by nonspecific electrostatic interactions. In contrast, the potassium and cesium ions specifically promote helix formation and aggregation (gelation) of furcellaran. The divalent calcium and magnesium ions are nonspecific, but more potent than the nonspecific monovalent ions in inducing helices. Anions differ in their capacity to stabilize the furcellaran helix in the sequence Cl? < NO < Br? < SCN? < I?. The iodide and thiocyanate anions impede aggregation and gel formation. 133Cs-nmr chemical shifts indicate specific binding of cesium ions to the furcellaran helix. Thus, with respect to its ion specificity and ion-binding properties, furcellaran, with 0.6 sulfate group per repeating disaccharide, resembles κ-carrageenan (1 sulfate/disaccharide) but differs from ι-carrageenan (2 sulfates/disaccharide). The conformational transition temperatures of furcellaran are, however, generally higher than those of κ-carrageenan under comparable conditions, and in mixtures of the two polysaccharides, separate transitions still occur, indicating that no mixed helices are formed. The observed ion sensitivity and cation-binding properties of furcellaran agree with predictions, by the PBCM, for a K-carrageenan with a reduced charge density.  相似文献   

2.
The binding effect of divalent cation Cu(2+) on the gelation process with a coil-helix transition in Cu(2+)/gellan aqueous solutions has been successfully elucidated by EPR, CD, and viscoelasticity measurements. Generally, Na-type gellan gum in aqueous solution can make gel when accompanied by an intrinsic coil-helix formation induced by hydrogen bonding between chains without any additional cations at T(ch)(-)(in) ( approximately 29 degrees C) with cooling temperature. An extrinsic coil-helix transition, induced by additional divalent cations in advance of the intrinsic sol-gel transition of gellan gum, is separately detected by CD measurement. The extrinsic coil-helix transition temperatures T(ch)(-)(ex) (>47 degrees C), which increased with the Cu(2+) concentration added, were nearly identical to the sol-gel transition temperature, T(sg), determined by the viscoelasticity measurement. Judging from the molar ellipticity by CD measurement and quantitative analysis of EPR spectra, it was elucidated that the helix forming process via divalent cations is composed of two steps ascribed to the different origins, i.e., a chemical binding effect via Cu(2+) ions in the initial stage and hydrogen bonds subsequently. Finally, we propose the coil-helix and the sol-gel transition mechanism initiated by the binding effect with the divalent cation, in which the partial chelate formation can cause local formation of helices and junction zones in the vicinity of the chelates at the initial stage of the process and stabilize the helices and the junction zones. On the other hand, the stabilized helices and junction zones can induce further formation and further stabilization of the Cu(2+)-gellan chelates. The mutual stabilization promotes the formation of three-dimensional network structure at the higher temperature than the intrinsic temperature for network formation.  相似文献   

3.
在自然盐碱生境下,通过测定不同月份土壤和马蔺体内主要阳离子Na+、K+、Ca2+、Mg2+的含量,研究了主要阳离子的吸收、转运变化及其在马蔺体内的分布.结果表明: 不同月份马蔺体内阳离子含量变动很大.在6月以后,随着马蔺的生长, Na+、K+、Ca2+和Mg2+4种离子在植物体内累积量逐渐增加.其中,根中Ca2+、Na+含量峰值出现在7月,分别为2.30%和0.51%,K+、Mg2+的含量峰值分别出现在9、10月,分别为0.27%和0.28%;叶片中Na+含量在7月达到最大值(0.57%);K+、Ca2+和Mg2+在8月分别达到1.30%、2.69%和0.47%.与Na+相比, 7、8月时马蔺对K+的选择吸收能力较低,但转运能力较强.马蔺对所测离子有很强的富集能力,各种离子在植物体内的含量都明显高于土壤背景值,且不同部位对离子的利用和累积能力不同,马蔺对各阳离子的累积主要集中在地上30 cm到地下40 cm范围内.马蔺地上部分平均单株K+、Na+、Ca2+和Mg2+含量分别是地下部分的9.11、4.07、0.98和2.27倍.  相似文献   

4.
Several rheological properties of aqueous alginate solution have been investigated in the absence and presence of four divalent cations: Ca, Cu, Mn, and Co. The concentration dependence of viscosity in alginate solution without addition of divalent cations shows a similar behavior to that found for other polysaccharides and synthetic polymers. We have found that for Ca(II) -added systems, sol-gel transition curves for different alginate concentrations can be superimposed over the concentration range of this study by using a normalized parameter. Shear-thinning behavior has been observed for all the alginate solutions by addition of four divalent cations over sol-gel transition process. The values of exponent b in the power-law relation $ {\eta \propto \dot \gamma }$ were found in the range of ?0.3 ~ ?0.6. It is suggested that a considerable portion of divalent cations remains in a cross-linking state even under shear flow. The results obtained from thixotropy measurements indicate the existence of a major framework formed in the gel-like and well-gelled samples, which can be maintained until shear rate (or shear stress) reaches a threshold value. © 1994 John Wiley & Sons, Inc.  相似文献   

5.
The selective permeability to monovalent metal cations, as well as the relationship between cation permeation and gating kinetics, was investigated for native tetrodotoxin-insensitive Na-channels in guinea pig ventricular myocytes using the whole-cell patch clamp technique. By the measurement of inward unidirectional currents and biionic reversal potentials, we demonstrate that the cardiac Na-channel is substantially permeable to all of the group Ia and IIIa cations tested, with the selectivity sequence Na(+) >/= Li(+) > Tl(+) > K(+) > Rb(+) > Cs(+). Current kinetics was little affected by the permeant cation species and concentrations tested (相似文献   

6.
We investigated a phenomenon of ultrasonic cleavage of DNA complexed with transition metal cations Ag(I), Cu(II) and Hg(II). We found the statistically significant dependence of relative intensity of cleavage on cation type and concentration. Each cation may cause two different types of distortion in the DNA double-helix depending on whether it binds to major or minor DNA groove. The intensity of ultrasonic cleavage decreases where the cation binds to the major DNA groove; the intensity of cleavage increases where the cation binds to the minor DNA groove and disturbs the hydrogen bonds of complementary base pairs or where it intercalates between bases. Both types of DNA distortion can affect the intensity of N?S intercon-version of deoxyribose.  相似文献   

7.
The chemical structure and the rheological behavior of the Klebsiella polysaccharide ATCC 12657 was studied and compared with data described in the literature and obtained for similar polysaccharides. The acetylated polysaccharide presents in solution a normal viscoelastic behavior with no evidence of an ordered conformation whatever the experimental conditions are. The deacetylated form can induce the formation of physical gels, in the presence of salt excess or ethanol. Microcalorimetry, optical rotation, and rheology experiments demonstrate that a thermally reversible and highly cooperative conformational transition occurs at the same temperature than a sol-gel transition. The melting of the gel and the conformational transition temperatures are dependent on the nature of cations and ionic concentration, whereas the gel strength is only influenced by polymer concentration.  相似文献   

8.
Metal complexes with peptide or pseudopeptide type ligands can serve as good model compounds for a deeper understanding of enzymatic catalysis, but ligands with a high selectivity for different transition metal cations are hard to find due to the rather flexible nature of peptides. Since such ligands would be the sine qua non condition for the synthesis of heterodinuclear peptide metal complexes with catalytic activity, the search for small, affine and selective metal chelating sequences is of interest. Using four different amino acids (His, Lys, Asp, Glu) a set of 16 pseudotripeptides of the common structure Bz-AS1-Sar-AS2-NH2 has been synthesized, purified and characterized by mass spectrometry and 1H-NMR. Their ability to form metal complexes has been investigated leading to short motifs capable of selectively binding only one or two transition metal cations with high affinity. As expected, the complexation of transition metal cations by pseudotripeptides is strongly dependent not only on the amino acid composition, but also on the sequence with regard to the stability of the resulting complexes, as well as the selectivity of the ligands towards Cu2+, Co2+, Ni2+, Zn2+ and Mn2+.  相似文献   

9.
Fibrin aggregation before sol-gel transition.   总被引:1,自引:0,他引:1       下载免费PDF全文
Fibrinogen solutions (concentrations 2 mg/ml, 0.15-M Tris-NaCl buffer, pH 7.4) were incubated at 20 degrees C with quantities of reptilase or thrombin that were so small that the polymerization process could be followed for several hours by means of static and dynamic light scattering. The scattered intensity and its correlation function were recorded at scattering angles between 30 degrees and 150 degrees. The measured data were compared with model calculations based on the Flory-Stockmayer distribution, which predicts a sol-gel phase transition. This distribution is characterized by a parameter, lambda, that indicates the extent of aggregation. lambda = 0 corresponds to the monomeric solution, and lambda = 1 indicates the sol-gel transition. Good agreement was found for monomeric units of 75-nm length aggregating (a) end-to-end in the early stage (0 less than or equal to lambda less than or equal to 0.3), and (b) in a staggered overlap pattern for the progressing polymerization (0.3 less than or equal to lambda less than 1). Before the gel point was reached, no systemic difference was observed between the data obtained after activation with thrombin which releases both fibrinopeptides A and B from fibrinogen, and reptilase, which exclusively releases the fibrinopeptides A. This confirms that the release of the fibrinopeptides A is the essential prerequisite for the aggregation process.  相似文献   

10.
Many RNAs, including the ribosome, RNase P, and the group II intron, explicitly require monovalent cations for activity in vitro. Although the necessity of monovalent cations for RNA function has been known for more than a quarter of a century, the characterization of specific monovalent metal sites within large RNAs has been elusive. Here we describe a biochemical approach to identify functionally important monovalent cations in nucleic acids. This method uses thallium (Tl+), a soft Lewis acid heavy metal cation with chemical properties similar to those of the physiological alkaline earth metal potassium (K+). Nucleotide analog interference mapping (NAIM) with the sulfur-substituted nucleotide 6-thioguanosine in combination with selective metal rescue of the interference with Tl+ provides a distinct biochemical signature for monovalent metal ion binding. This approach has identified a K+ binding site within the P4-P6 domain of the Tetrahymena group I intron that is also present within the X-ray crystal structure. The technique also predicted a similar binding site within the Azoarcus group I intron where the structure is not known. The approach is applicable to any RNA molecule that can be transcribed in vitro and whose function can be assayed.  相似文献   

11.
H J Hinz  W Haar  T Ackermann 《Biopolymers》1970,9(8):923-936
The enthalpies of the helix-coil transitions of the ordered polynucleotide systems of poly(inosinic acid)–poly(cytidylic acid) [poly(I + C)], (helical duplex), and of poly (inosinic acid) [poly(I + I + I)], (proposed secondary structure: a triple-stranded helical complex), were determined by using an adiabatic twin-vessel differential calorimeter. Measuring the temperature course of the heat capacity of the aqueous polymer solutions, the enthalpy values for the dissociation of the helical duplex poly (I + C) and the three-stranded helical complex poly(I + 1 + 1), respectively, were obtained by evaluating the additional heat capacity involved in the conformational change of the polynucleotide system in the transition range. The ΔH values of the helix-coil transition of poly (I + C) resulting from the analysis of the calorimetric measurements vary between the limits 6.5 ± 0.4 kcal/mole (I + C) and 8.4 ± 0.4 kcal/mole (I + C). depending on the variation of the cation concentration ranging from 0.063 mole cations kg H2O to 1.003 mole cations/kg H2O. The calorimetric investigation of an aqueous poly I solution (cation concentration 1.0 mole/kg H2O) yielded the enthalpy value ΔH = 1.9 ± 0.4 kcal/mole (I), a result which has been interpreted qualitatively following current models of inter- and intramolecular forces of biologically significant macromolecules. Additional information on the transition behavior of poly(I+ C)Was obtained by ultraviolet and infrared absorption measurements.  相似文献   

12.
ATP-gated P2X(2) channels undergo activation-dependent permeability increases as they proceed from the selective I(1) state to the I(2) state that is readily permeable to organic cations. There are two main models about how permeability changes may occur. The first proposes that permeability change-competent P2X channels are clustered or redistribute to form such regions in response to ATP. The second proposes that permeability changes occur because of an intrinsic conformational change in P2X channels. In the present study we experimentally tested these views with total internal reflection fluorescence microscopy, electrophysiology, and mutational perturbation analysis. We found no evidence for clusters of P2X(2) channels within the plasma membrane or for cluster formation in response to ATP, suggesting that channel clustering is not an obligatory requirement for permeability changes. We next sought to identify determinants of putative intrinsic conformational changes in P2X(2) channels by mapping the transmembrane domain regions involved in the transition from the relatively selective I(1) state to the dilated I(2) state. Initial channel opening to the I(1) state was only weakly affected by Ala substitutions, whereas dramatic effects were observed for the higher permeability I(2) state. Ten residues appeared to perturb only the I(1)-I(2) transition (Phe(31), Arg(34), Gln(37), Lys(53), Ile(328), Ile(332), Ser(340), Gly(342), Trp(350), Leu(352)). The data favor the hypothesis that permeability changes occur because of permissive motions at the interface between first and second transmembrane domains of neighboring subunits in pre-existing P2X(2) channels.  相似文献   

13.
The vibrational Raman spectra of both pure 1-alpha-dimyristoylphosphatidic acid (DMPA) liposomes and DMPA multilayers reconstituted with ferricytochrome c at pH 7 and pH 4, with either sodium or calcium as the cation, are reported as a function of temperature. Multilayers composed of a 1:1 mol ratio DMPA and dimyristoylphosphatidylcholine with perdeuterated acyl chains (DMPC-d54) have also been reconstituted with approximately 10(-4) M ferricytochrome c for Raman spectroscopic observation. Total integrated band intensities and relative peak height intensity ratios, two spectral Raman scattering parameters used to characterize bilayer properties, are sensitive to the presence of both ferricytochrome c and the cation in the reconstituted liposomes. Temperature profiles, derived from the various Raman intensity parameters for the 3,100-2,800 cm-1 lipid acyl chain C-H stretching mode region specifically reflect bilayer perturbations due to the interactions of ferricytochrome c. At pH 4 the calcium DMPA multilamellar gel to liquid crystalline phase transition temperatures Tm, defined by either the C-H stretching mode I2850/I2880 and I2935/I2880 peak height intensity ratios, are 58.5 +/- 0.5 degrees C and 60.0 +/- 0.3 degrees C, respectively. This difference in Tm's resolves the phase transition process into first an expansion of the lipid lattice and then a melting of the lipid acyl chains. At pH 7 the calcium DMPA liposomes show no distinct phase transition characteristics below 75 degrees C. For sodium DMPA liposomes reconstituted with ferricytochrome c at either pH 4.0 or pH 7.0, spontaneous Raman spectra show altered lipid structures at temperatures above 40 degrees C. Resonance Raman spectra indicate that ferricytochrome c reconstituted in either calcium or sodium DMPA liposomes changes irreversibly above Tm. For either the binary lipid or ternary lipid-protein systems reconstituted with DMPC-d54, linewidth parameters of the DMPC-d54 acyl chain CD2 symmetric stretching modes at 2,103 cm-1 provide a sensitive measure of the conformational and dynamic properties of the perdeuterated lipid component, while the 3,000 cm-1 C-H spectral region reflects the bilayer characteristics of the DMPA species in the complex. Although calcium clearly induces a lateral phase separation in the DMPA/DMPC-d54 system at pH 7.5 (Kouaouci, R., J.R. Silvius, I. Grah, and M. Pezolet. 1985. Biochemistry. 24:7132-7140), no distinct lateral segregation of the lipid components is observed in the mixed DMPA/DMPC-d54 lipid system in the presence of either ferricytochrome c or the sodium and calcium cations at pH 4.0.(ABSTRACT TRUNCATED AT 400 WORDS)  相似文献   

14.
Many ion channels have wide entrances that serve as transition zones to the more selective narrow region of the pore. Here some physical features of these vestibules are explored. They are considered to have a defined size, funnel shape, and net-negative charge. Ion size, ionic screening of the negatively charged residues, cation binding, and blockage of current are analyzed to determine how the vestibules influence transport. These properties are coupled to an Eyring rate theory model for the narrow length of the pore. The results include the following: Wide vestibules allow the pore to have a short narrow region. Therefore, ions encounter a shorter length of restricted diffusion, and the channel conductance can be greater. The potential produced by the net-negative charge in the vestibules attracts cations into the pore. Since this potential varies with electrolyte concentration, the conductance measured at low electrolyte concentrations is larger than expected from measurements at high concentrations. Net charge inside the vestibules creates a local potential that confers some cation vs. anion, and divalent vs. monovalent selectivity. Large cations are less effective at screening (diminishing) the net-charge potential because they cannot enter the pore as well as small cations. Therefore, at an equivalent bulk concentration the attractive negative potential is larger, which causes large cations to saturate sites in the pore at lower concentrations. Small amounts of large or divalent cations can lead to misinterpretation of the permeation properties of a small monovalent cation.  相似文献   

15.
The effects of divalent cations on the E-4031-sensitive repolarization current (I(Kr)) were studied in single ventricular myocytes isolated from rabbit hearts. One group of divalent cations (Cd2+, Ni2+, Co2+, and Mn2+) produced a rightward shift of the I(Kr) activation curve along the voltage axis, increased the maximum I(Kr) amplitude (i.e., relieved the apparent inward rectification of the channel), and accelerated I(Kr) tail current kinetics. Another group (Ca2+, Mg2+ and Sr2+) had relatively little effect on I(Kr). The only divalent cation that blocked I(Kr) was Zn2+ (0.1-1 mM). Under steady-state conditions, Ba2+ caused a substantial block of I(K1) as previously reported. However, block by Ba2+ was time dependent, which precluded a study of Ba2+ effects on I(Kr). We conclude that the various effects of the divalent cations can be attributed to interactions with distinct sites associated with the rectification and/or inactivation mechanism of the channel.  相似文献   

16.
The effect of hexachlorocyclohexane (all isomers) on the thermal transition properties of phospholipid liposomes was determined by Raman spectroscopy. Raman spectra of liposomes with and without the presence of hexachlorocyclohexanes were recorded in the C-H stretching region which shows three major bands around 2850, 2880 and 2930 cm-1. Thermal transition properties were estimated from plots of I2880/I2850 and or I2930/I2850 vs. temperature, where I represents the intensity of the respective band. Our data on phospholipid liposomes reveal that delta- and gamma-hexachlorocyclohexanes drastically reduce and broaden the main thermal transitions of phospholipids at toxic level concentrations. These effects are more pronounced in liposomes containing 18 or more carbon atom long acyl chains. Alpha- and beta-isomers at similar concentrations show a minimum effect on the thermal transition properties of phospholipids. Raman analysis of phospholipid liposomes containing melittin, interestingly, reveal that the delta-isomer unlike the gamma-isomer strongly alters the transition properties of boundary lipids. These data suggest that the effect of hexachlorocyclohexanes on the thermal transition properties of membranes is stereo specific and that the delta-isomer preferably disrupts the lipid-protein domains. Results are explained on the basis of the dynamic flexibility owing to the equatorial and axial chlorine atoms of various hexachlorocyclohexane isomers.  相似文献   

17.
Effect of inorganic cations on phase transitions.   总被引:4,自引:0,他引:4  
The effect of protons and cations on the crystal (gel)-to-liquid crystal transition temperature Tm of isoelectric and negatively charged phospholipids are summarized. The general trends emerging are as follows: Tm depends on the state of ionization of the phospholipid in that Tm-vs-pH-curves parallel the titration curve of the phospholipid. Protonation of phospholipids causes Tm to increase, deprotonation or ionization has the opposite effect. The effects of cations on the Tm of phospholipids may be grouped into non-specific and specific effects. Unspecific effects of cations such as the screening of negative charges of the phospholipid polar group are qualitatively similar to protonation: Tm increases, in the order monovalent less than divalent less than trivalent cations and the effects on negatively charged phospholipids are larger than those on isoelectric phospholipids. Unspecific, electrostatic effects on Tm are reasonably well accounted for by the Gouy-Chapman theory. If, however, specific binding comes into play and/or electrostatic effects are accompanied by changes in phospholipid structure, simple, electrostatic theories fail to explain the observed changes in Tm. The crystal (gel)-to-liquid crystal transition is also a function of the degree of hydration: Tm generally decreases with increasing hydration reaching a plateau in excess H2O. In addition to screening of electric charges, ions may exert yet another non-specific effect: ions may affect Tm indirectly by competing with the phospholipid polar group for water of hydration. This indirect effect plays a role at high ionic strength and/or at low hydration of the phospholipid. Specific binding of cations to negatively charged phospholipids can lead to tight associations of the metal ion with the lipid polar group. Isothermal crystallization of the phospholipid bilayer is induced that is accompanied by a total or partial loss of water of hydration resulting in a marked increase in Tm. For instance, in crystalline Ca2(+)-phosphatidylserine complexes Tm is increased by more than 100 degrees C.  相似文献   

18.
We have examined the Raman scattering due to CH stretching vibrations, as well as to v(-C=C-) and v(=C-C=) of beta-carotene, of liposomes composed of phosphatidylcholine (egg, dimyristoyl, dipalmitoyl) +/- cholesterol, beta-carotene or melittin in the temperature range of -10 degrees C to 45 degrees C. (2) Plots vs. temperature of the intensities of the 2885 cm-1 and 2930 cm-1 CH stretching bands relative to the intensity of the thermally stable 2850 cm-1 band, i.e. the I2885/I2850 and I2930/I2850 ratios, reveal a sharp discontinuity in cholesterol-free phosphatidylcholine liposomes; this coincides with the gel leads to liquid-crystal transition temperature of the fatty acyl chains. In cholesterol/phosphatudylcholine liposomes the change in I2885/I2850 occurs over a very broad temperature range and I2930/I2850 remains stable. (3) I1527/I1158, i.e. the intensity of v(-C=C-) relative to that of v(=C-C-) in beta-carotene/phosphatidylcholine liposomes, changes discontinuously at the gel leads to liquid-crystal transition temperature. The values above the transition temperature approach those of the carotenoid in organic solution. (4) The transitions reported in I2885/I2850 for phosphatidylcholine/melittin liposomes (25-56; 1, M/M) are shifted to much higher temperatures than observed in phosphatidylcholine liposomes. In the case of dimyristoyl phosphatidylcholine/melittin the changes in I2930/I2850 also occurs at a higher temperature (28 degrees C) than without melittin (21 degrees C), but the temperature shift is less than the +13 degrees C observed for I2885/I2850. It appears that the apolar moiety of melittin organizes phospholipids adjacent to and more remote from the peptide moiety, to form complexes with an elevated lipid transition temperature. The effect of the peptide moiety is greater on the methylene segments (I2885/I2850) than on the methyl termini (I2930/I2850).  相似文献   

19.
RNA polymerase I was isolated from parsley cells grown in suspension culture and from soybean hypocotyls. Kinetic studies of the enzyme revealed that RNA polymerase I is an allosteric regulated enzyme. The enzyme activity was influenced by nucleoside triphosphates (NTP) and divalent cations. NTP exceeding a 1:1 ratio of these two components acted as allosteric inhibitors, contrary to free divalent cations, which had promotive effects on the RNA polymerase I. Furthermore, isolated nuclei from parsley exhibited a powerful nucleoside triphosphatase (NTPase) activity. Contrary to RNA polymerase I, this enzyme was stimulated by NTP exceeding the 1:1 ratio of NTP and divalent cations. Free divalent cations had an inhibitory effect. Assuming that a causal connection of these two processes does exist, a possible role of this NTPase would be the control of NTP pools in relation to divalent cations and thus regulating RNA synthesis.  相似文献   

20.
The problems of interdisciplinary interests--applying methods of graph theory to sol-gel patterns and to cytoskeleton--are discussed. The importance of sol-gel transition phenomena in living cells and the possibility of periodic sol-gel transition phenomena are briefly reviewed. Representing patterns by graphs and using graph probabilistic representation for calculating structure-property relationships are discussed and applied to sol-gel transition patterns.  相似文献   

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