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1.
The effects of organic solvents on the 31P-mr chemical shifts of various phosphate diesters have been investigated in water and mixed-organic solvent systems. The addition of organic solvents to cyclic phosphates and to diethyl phosphate causes large upfield shifts of the phosphorus resonance which are attributed to solvent-induced changes in the local hydration of the phosphodiester group. This is consistent with the fact that there is an inverse correlation between the hydrogen-bond-donating ability of the solvents and the magnitude of the shifts they induce. Other possible interpretations, such as solvent-induced ion pairing and solvent-induced conformational changes, appear to be eliminated. Fourier-transform ir study of the cyclic nucletides reveals that there are also large solvent-induced shifts in the frequency of the antisymmetric OPO stretching frequency, and a comparison of the two types of measurements indicates that there is a linear correlation between shifts observed in the ir and in the 31P-nmr spectra. With UpU, the solvent-induced 31P-nmr shifts are ~3 times smaller than those observed with the cyclic phosphates and the solvent-induced shift of the OPO band is reduced (factor of ~1.7) as compared with the cyclic phosphates. With the single-stranded polynuclotides, poly(C) and poly(U), the solvent-induced shifts in both the nmr and ir are quite small (~0.1 ppm and ~1 cm?1). The very small solvent effects observed with poly(U) and poly(C) are attributed to a combination of steric effects and a polyelectrolyte effect which maintains a high density of counterions with waters of hydration in the vicinity of the charged backbone and makes the phosphates much less susceptible to solvent-induced changes in hydration. 相似文献
2.
T Weidlich S M Lindsay Q Rui A Rupprecht W L Peticolas G A Thomas 《Journal of biomolecular structure & dynamics》1990,8(1):139-171
We have obtained low frequency (less than 200 cm-1) Raman spectra of calf-thymus DNA and poly(rI).poly(rC) as a function of water content and counterion species and of d(GGTATACC)2 and d(CGCGAATTCGCG)2 crystals. We have found that the Raman scattering from water in the first and second hydration shells does not contribute directly to the Raman spectra of DNA. We have determined the number of strong Raman active modes by comparing spectra for different sample orientations and polarizations and by obtaining fits to the spectra. We have found at least five Raman active modes in the spectra of A- and B-DNA. The frequencies of the modes above 40 cm-1 do not vary with counterion species, and there are only relatively small changes upon hydration. These modes are, therefore, almost completely internal. The mode near 34 cm-1 in A-DNA is mostly internal, whereas the mode near 25 cm-1 is dominated by interhelical interactions. The observed intensity changes upon dehydration were found to be due to the decrease in interhelical distance. Polymer length appears to play a role in the lowest frequency modes. 相似文献
3.
Zhang Q Andrew Chan KL Zhang G Gillece T Senak L Moore DJ Mendelsohn R Flach CR 《Biopolymers》2011,95(9):607-615
Water is an integral part of collagen's triple helical and higher order structure. Studies of model triple helical peptides have revealed the presence of repetitive intrachain, interchain, and intermolecular water bridges (Bella et al., Structure 1995, 15, 893-906). In addition, an extended cylinder of hydration is thought to be responsible for collagen fiber assembly. Confocal Raman spectroscopy and dynamic vapor sorption (DVS) measurements of human Type I collagen and pigskin dermis were performed to probe relative humidity (RH)-dependent differences in the nature and level of collagen hydration. Raman spectra were also acquired as a function of time for both Type I collagen and pigskin dermis samples upon exchange of a 100% RH H(2) O to deuterium oxide (D(2) O) environment. Alterations in Amide I and III modes were consistent with anticipated changes in hydrogen bonding strength as RH increased and upon H → D exchange. Of note is the identification of a Raman spectral marker (band at 938 cm(-1) ) which appears to be sensitive to alterations in collagen-bound water. Analysis of DVS isotherms provided a quantitative measure of adsorbed and absorbed water vapor consistent with the Raman results. The development of a Raman spectral marker of collagen hydration in intact tissue is relevant to diverse fields of study ranging from the evaluation of therapeutics for wound healing to hydration of aging skin. 相似文献
4.
Kuntz and Kauzmann have argued that dehydrating a protein results in conformational changes. In contrast, Rupleyet al. have developed a hydration model which involves no significant change in conformation; the onset of enzyme activity in hen
egg-white lysozyme at hydration values of about 0.2 g water/g protein they ascribe rather to a solvation effect. Using a direct
difference infra-red technique we can follow specific hydration events as water is added to a dry protein. Conformational
studies of lysozyme using laser Raman spectroscopy indicate changes in conformation with hydration that are complete just
before measurable activity is found. Parallel nuclear magnetic resonance measurements of exchangeability of the main chain
amide hydrogens, as a function of hydration from near dryness, suggest a hydration-related increase in conformational flexibility
which occurs before-and is probably necessary for-the Raman-detected conformational changes. Very recent inelastic neutron
scattering measurements provides direct evidence of a flexibility change induced by hydration, which is apparently necessary
before the enzyme can achieve adequate flexibility for it to begin to function. 相似文献
5.
A critical review of the nucleosidic vibration modes appearing in the 800-500-cm-1 spectral region, based on new harmonic dynamics calculations 总被引:1,自引:0,他引:1
On the basis of a harmonic dynamics calculation, it is shown that in the 800–500-cm?1 spectral region of DNA vibrational spectra, the characteristic Raman peaks and ir bands do not arise from the same nucleosidic motions. The Raman spectra involve mainly the ring-breathing modes of nucleic bases while the ir spectra reveal essentially their out-of-plane vibrations. Moreover, the calculated results show the splitting of the guanine- and adenine-residue breathing modes upon their coupling with the sugar-pucker motions. This fact is in agreement with the poly[d(G-C)] and poly[d(A-T)] Raman spectra. 相似文献
6.
Thermal properties of water in myoglobin crystals and solutions at subzero temperatures. 总被引:8,自引:7,他引:8
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The water of hydration in myoglobin crystals and solutions was studied at subzero temperatures by calorimetry and infrared spectroscopy (ir). For comparison we also investigated glycine, DL-alanine and DL-valine solutions. The hydration water remains amorphous at low temperatures. We find a broad glass transition between 180 and 270 K depending on the degree of hydration. The ice component shows a noncolligative melting point depression that is attributed to a finite conformational flexibility. The ir spectrum and the specific heat of water in myoglobin crystals was determined for the first time between 180 and 290 K. The glass transition in crystals is qualitatively similar to what is found in amorphous samples at the same water content. These data are compared with M?ssbauer experiments and dielectric relaxation of water in myoglobin crystals. The similar temperature dependencies suggest a cross correlation between structural fluctuations and the thermal motion of crystal water. A hydrogen bond network model is proposed to explain these features. The essential ingredients are cooperativity and a distribution of hydrogen-bonded clusters. 相似文献
7.
Y A Lazarev B A Grishkovsky T B Khromova A V Lazareva V S Grechishko 《Biopolymers》1992,32(2):189-195
The ir amide bands of the triple-helical polytripeptides and collagens upon hydration of films are investigated. On the basis of our assignment of the amide I components, the formation of hydrogen bonds between the peptide backbone and structural water is studied. The C1O1--HOH hydrogen bonds are found more ordered than the C3O3--HOH hydrogen bonds. The specific incorporation of water in the triple helix is followed by multistep conformational changes and by increasing of the interpeptide hydrogen-bond strength. The formation of the polypeptide hydrate structure depending on the amino acid composition and the chain length is examined. 相似文献
8.
Scott A. Lee Nong-Jian Tao Allan Rupprecht 《Journal of biomolecular structure & dynamics》2013,31(11):1337-1342
Raman spectroscopy is used to probe the nature of the hydrogen bonds which hold the water of hydration to DNA. The ~ 3450?cm?1 molecular O–H stretching mode shows that the first six water molecules per base pair of the primary hydration shell are very strongly bound to the DNA. The observed shift in the peak position of this mode permits a determination of the length of the hydrogen bonds for these water molecules. These hydrogen bonds appear to be about 0.3?Å shorter than the hydrogen bonds in bulk water. The linewidth of this mode shows no significant changes above water contents of about 15 water molecules per base pair. This technique of using a vibrational spectroscopy to obtain structural information about the hydration shells of DNA could be used to study the hydration shells of other biomolecules. 相似文献
9.
Direct difference ir spectra are presented as a function of hydration for lysozyme and α-lactalbumin, and detailed sequential hydration molecular events identified. Despite the strong sequence homology between the two proteins, and their expected conformational similarity, the hydration behaviour of the polar groups is different for the two proteins. Using a Hill-type analysis, we conclude that the acid groups ionize and hydrate rapidly and noncooperatively in both proteins, consistent with the known (lysozyme) and postulated (α-lactalbumin) surface chemistry. The polar group hydration shows a clear cooperativity, which is quantitatively different in the two proteins. Complementary work suggests this cooperativity relates to a hydration-induced “loosening up” of the lysozyme conformation at about 55 mol water/mol protein. α-Lactalbumin appears to “open up” more easily for hydration than does lysozyme, consistent with its lower stability against thermal and acid denaturation. 相似文献
10.
By a combined gravimetric and ir technique, spectra of protein films are recorded during sorption isotherms at constant water content h (mg D2O/mg dry protein) in the range 0 les; h ? 0.35 at 27 and 38°C. Computer-aided differential analysis shows the effect of progressive hydration on some significant sites of the protein such as the ionizable acidic side chains and the backbone amide carbonyls, as well as the spectrum of the adsorbed water itself. In order to derive thermodynamic properties of these sites, the measured sorption isotherm is decomposed in terms of a model which postulates the existence of two classes of primary sorption sites only, and these two contributions are independently checked by the ir data. The free energy of binding of the strong and weak binding sites is found to be 2.0 ± 0.2 and 0.40 ± 0.1 kcal/mol, respectively. A water-induced transition region is clearly detected in all the observed properties at 0.06 < h < 0.10 at 38°C and is shown to be due to changes involving both the structure of the absorbed water and the coverage of the absorption sites. A detailed picture of the hydration events is offered, and the relevance of these findings to protein dynamics is discussed. 相似文献
11.
Low-frequency Raman spectra of lysozyme crystals and oriented DNA films: dynamics of crystal water.
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We observed low-frequency Raman spectra of tetragonal lysozyme crystals and DNA films, with varying water content of the samples. The spectra are fitted well by sums of relaxation modes and damped harmonic oscillators in the region from approximately 1 cm(-1) to 250 cm(-1). The relaxation modes are due to crystal water, and the distribution of relaxation times is determined. In wet samples, the relaxation time of a small part of the water molecules is a little longer than that of bulk water. The relaxation time of a considerable part of the crystal water, which belongs mainly to the secondary hydration shell, is an order of magnitude longer than that of bulk water. Furthermore, the relaxation time of some water molecules in the primary hydration shell of semidry samples is shorter than we expected. Thus we have shown that low-frequency Raman measurements combined with properly oriented samples can give specific information on the dynamics of hydration water in the ps range. On the other hand, we concluded, based on polarized Raman spectra of lysozyme crystals, that the damped oscillators correspond to essentially intramolecular vibrational modes. 相似文献
12.
The rather thin outermost layer of the mammalian skin, stratum corneum (SC), is a complex biomembrane which separates the water rich inside of the body from the dry outside. The skin surface can be exposed to rather extreme variations in ambient conditions (e.g. water activity, temperature and pH), with potential effects on the barrier function. Increased understanding of how the barrier is affected by such changes is highly relevant for regulation of transdermal uptake of exogenous chemicals. In the present study we investigate the effect of hydration and the use of a well-known humectant, urea, on skin barrier ultrastructure by means of confocal Raman microspectroscopy. We also perform dynamic vapor sorption (DVS) microbalance measurements to examine the water uptake capacity of SC pretreated with urea. Based on novel Raman images, constructed from 2D spectral maps, we can distinguish large water inclusions within the skin membrane exceeding the size of fully hydrated corneocytes. We show that these inclusions contain water with spectral properties similar to that of bulk water. The results furthermore show that the ambient water activity has an important impact on the formation of these water inclusions as well as on the hydration profile across the membrane. Urea significantly increases the water uptake when present in skin, as compared to skin without urea, and it promotes formation of larger water inclusions in the tissue. The results confirm that urea can be used as a humectant to increase skin hydration. 相似文献
13.
Dieter Naumann Harald Labischinski Wolfgang Rnspeck Gerhard Barnickel Hans Bradaczek 《Biopolymers》1987,26(6):795-817
The synthetic, zwitterionic bacterial cell wall peptides—D -Gluγ-L-Lys, D -Gluγ-L-Lys-D -Ala, D -Gluγ-L-Lys-D -Ala-D -Ala, and L-Ala-D -Gluγ-L-Lys-D -Ala-D -Ala—have been investigated in the crystalline and aqueous solution state applying ir and Raman spectroscopy. Additionally, aqueous solutions of the tetra- and pentapeptide have been investigated by CD spectroscopic techniques. Apart from the dipeptide, whose spectral features were dominated by end-group vibrations, the corresponding ir and Raman active bands of the crystalline peptides in the amide and skeletal regions were found at similar wave numbers, thus suggesting an analogous three-dimensional structure of these compounds. Dominant amide A, I, II, and III bands near 3275, 1630, 1540, and 1220–1250 cm?1, respectively, in the ir are interpreted in favor of an intermolecularly hydrogen-bonded, β-like structure. The absence of any amide components near 1680–1690 cm?1, together with the presence of strong amide bands near 1630 cm?1, and weak bands near 1660 cm?1 in the ir, which, conversely, were found in the Raman spectra as weak and strong bands, but at corresponding wave numbers, is taken as strong evidence for the presence of the unusual, parallel-arranged β-structure. On the basis of comparative theoretical considerations, a parallel-arranged, “β-type ring” conformation [P. De Santis, S. Morosetti, and R. Rizzo (1974) Macromolecules 7 , 52–58] is hypothesized. The solubilized peptides exhibited distinct similarities with their crystalline counterparts in respect to frequency values and relative intensities of the corresponding ir and Raman-active amide I/I′ components, and of some Raman bands in the skeletal region. This is interpreted in terms of residual short-range order, persisting even in aqueous solution. We concluded that the peptides show a strong propensity to form hydrated, strongly associated aggregates in water. On the basis of amide I/I′ band positions, stable, intramolecular interactions via the amide groups are discarded for the solubilized peptides. Complementarily, the CD data obtained suggest the presence of weakly bent, “open-turn”-like structures for the tetra- and pentapeptide in aqueous solution. 相似文献
14.
Response of the local heme environment of (carbonmonoxy)hemoglobin to protein dehydration 总被引:1,自引:0,他引:1
The effects of protein dehydration upon the equilibrium and dynamic properties of the heme active site in human hemoglobin (HbA) have been probed by resonance Raman scattering. Spectra of equilibrium carbonmonoxy-HbA and the photolytic heme transient species generated within 10 ns of ligand photolysis have been obtained from thin films of protein in various stages of dehydration. These data provide detailed information concerning the response of the heme and its bonding interactions with both the proximal histidine and carbon monoxide as a function of protein hydration. For protein hydration levels of 0.4-1.0 g of H2O/g of protein, our results indicate that the C = O stretching mode of carbonmonoxy-HbA is dramatically affected by protein hydration levels, thus corroborating the infrared results of Brown et al. [Brown, W. E., Sutcliffe, J. W., & Pulsinelli, P. D. (1983) Biochemistry 22, 2914-2923]. However, we find that both heme skeletal modes and the Fe-C bond strength are largely insensitive to dehydration. Moreover, the proximal pocket geometry (as reflected in the behavior of the Fe-proximal histidine stretching mode) immediately following ligand photolysis was found to be very similar to that of R-state solution hemoglobin. At protein hydration levels below the theoretical monolayer limit, small changes in the resonance Raman spectra of both equilibrium HbCO and the transient heme species generated subsequent to ligand photolysis are detected. These include broadening of the Fe-C stretching mode in equilibrium HbCO and a small shift to lower frequency of the Fe-His mode in the photolytic transient species.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
15.
The Raman and ir spectra of a murine IgG2a monoclonal antibody molecule are reported. In accordance with previous studies on immunoglobulins, the secondary structure is predominantly of the beta-sheet type. The low frequency region of the Raman spectrum was also analyzed in detail. A structured band with two maxima near 43 and 94 cm(-1) was observed. This band has been attributed to vibrations of elastic body. The results are interpreted using a simple elastic model. 相似文献
16.
To explore the interaction potential of phospholipids, we have studied the hydration of diacyl phosphatidylcholine (PC) and methylphosphocholine (MePC), a pertinent model compound, by ir spectroscopy. Related ab initio Hartree-Fock calculations were performed for MePC. Water is considered ideal as a relevant probe molecule. Spectroscopic data for MePC reveal a strong influence of bound hydration water not only on the phosphate groups but also onto the putatively apolar CH(n) groups. The same could be demonstrated for deuterated dimyristoyl PC taken as a "complete" lipid molecule: both headgroup methyl and methylene moieties are gradually, but remarkably affected by hydration, as evidenced by strong wavenumber upshifts of C-H stretching vibration bands. These findings may originate in directed interactions of the CH(n) groups with bound water molecules, but hydration-driven conformational changes of PC headgroups could also occur. The results of the ab initio calculations rationalize the first explanation by predicting a substantial contribution of specific C-H...OH(2) interactions, mainly characterized by a dramatic loss of electron density of the sigma* antibonding molecular orbitals of C-H bonds. Hence, the propensity of the lipid headgroup methyl and methylene groups to act as donor sites in hydrogen bonding must no longer be ignored when considering the interaction potential of PCs. 相似文献
17.
Infrared and Raman spectra of the Mg2+ salt of poly(U) in D2O were recorded in the 1600-1800 cm?1 region and between 1 and 20C. The ir spectra showed a melting curve similar to the uv melting curves with a temperature of transition of about 6.5°C. This spectral change is assumed to be associated with the formation of the secondary structure of Mg2+-poly(U) in D2O at this temperature. Three double-helical and two triple-helical structures were used as inputs to compute the normal modes of vibration. A double-helical structure was found to give the best agreement with the observations. Knowledge of the C=0 eigenvectors, and of the expression for transition probability from quantum mechanics, was used to explain the so far unanswered question of H. T. Miles [(1964) Proc. Natl. Acad. Sci. USA 51, 1104–1109; (1980) Biomolecular Structure, Conformation, Function and Evolution, Pergamon, Oxford, pp. 251–264] as to why there is an increase in the ir vibrational wave number of a carbonyl band when that group is H-bonded to another polynucleotide chain in a helix. Such considerations also explain why a predicted band at about 1648 cm?1 is not to be seen in the ir spectra but is present in the Raman spectra. The model incorporating the C?O transition dipole-dipole coupling interaction is able to explain also the observed higher intensity of the higher wave-number ir band. The experimental results demonstrate that the complete picture of vibrational dynamics of Mg2+-poly(U) in D2O is obtained only by looking simultaneously at ir and Raman spectra and not at only one of them. Weak ir bands were found to be as useful as the strong ones in understanding structure and vibrational dynamics. On the bases of our ir and Raman spectra, of the normal-mode analyses, and of the literature data, it is concluded that Mg2+-poly(U) in D2O is present in a double-helical structure at temperatures below the temperature of transition, whereby the uracil residues are paired according to arrangement (a) (see Fig. 1). This structure is rodlike and arises by refolding of one poly(U) chain. The computations show that no normal mode is associated with a single C?O group vibration; all C?O group vibrations are heavily mixed motions of various C?O groups. 相似文献
18.
Wool fibers are comprised of proteins known as α-keratins and have a complex morphological structure. The major components of this structure, the cuticle and cortical cells, differ in the conformations of their peptide chains as well as their amino acid compositions. High quality Fourier transform Raman spectra of cortical and cuticle cells isolated from fine Merino wool fibers have been obtained. Raman spectroscopy has been shown to be sensitive to the differences in both secondary structure and amino acid composition. The cortical cells were found to be higher in α-helical content as compared to the cuticle cells, which had an increased disordered content. Specific information, consistent with amino acid analysis results, regarding cystine, tyrosine, tryptophan, and phenylalanine residues, were obtained for both the cortical and cuticle cells. In addition, the Raman spectra provided information about free thiol groups, amino acids residues with amide group side chains, and residues with protonated carboxyl group side chains. Middle ir transmission spectra of these isolated cells were also obtained. In comparison to the Raman data, the middle ir spectra were found to be not as rich in information. © 1997 John Wiley & Sons, Inc. Biopoly 42: 7–17, 1997 相似文献
19.
Raman spectral studies of nucleic acids. XI. Conformations of yeast tRNAPhe and E. coli ribosomal RNA in aqueous solution and in the solid state 总被引:2,自引:0,他引:2
Laser-excited Raman spectra of tRNAPhe from yeast and of fractionated 16S and 23S rRNA from E. coli are reported for samples in aqueous solution and in the solid state. The Raman scattering spectrum of each RNA is not significantly altered by the change from an aqueous to a solid environment and displays the same characteristic frequencies and intensities associated with ordered polyribonucleotide structures. Unlike DNA, the backbone conformation of RNA thus appears to be largely insensitive to gross changes in the degree of hydration. Raman scattering from the phosphate group vibrations of aqueous tRNAyeastPhe is qualitatively and quantitatively the same as obtained from previously studied tRNA's and is indicative of a highly ordered conformational structure in which some 85% of the nucleotide residues are in ordered configurations. The major differences observed between spectra of tRNA and rRNA are attributed to differences in base composition of these RNA's. 相似文献
20.
In-situ confocal Raman spectroscopy combined with relative humidity (RH) control technique was used to study the sequential dehydration process of insulin crystals. By gradually decreasing the ambient RH of the insulin crystal, the content of the hydration water in the crystal was quantitatively controlled. Tyrosine (Tyr) residues were very sensitive to the micro-environmental changes, and four Raman features 828cm(-1), 852cm(-1), 1174cm(-1) and 1206cm(-1) of Tyr were employed to monitor the dehydration process. Taking advantage of the ratios I(852)/I(828) at different RH values, the mole fractions of the 'exposed' and 'buried' Tyr residues were estimated. Moreover, using the ratio I(1174)/I(1206) as an indicator of the dehydration process, three RH regions were discriminated. This is believed to imply that different types of the hydration water were lost step by step, i.e. firstly the 'second-layer' and 'first-layer' classes, then the 'contact' class, and finally, the 'inside' class. In addition, the profile of the amide I band was observed to gradually change with RH. By band fitting of the amide I region, changes in secondary structure were quantitatively determined. And the results showed that nearly 17% of α-helix converted into β-sheet with RH decreasing from 92% to 2%. 相似文献