Rotovibrational states of the water molecule on the sun

The infrared spectrum of water observed in sunspots is complex and dense, with bands separated by approximately 0.01 cm^?1. For top asymmetrical molecules, there is no theoretical approach that allows for the calculation of rotovibrational energy with such precision. Experimentally derived rotovibracional energy levels of water at high temperatures combined with variational calculations have been used for the band assignments. These energy levels are employed to refine the analysis of a small portion of the infrared absorption spectrum. Such procedure has allowed for the identification of additional 55 bands to the 70 already identified as rotovibrational transitions of the water molecule. Our new assignments, which include pure and cross transitions, offer additional evidence of the existence of water on the sun, but above all they illustrate the complexity of the solar spectrum that involves states with higher levels of rotational excitation. Given the conditions on the sun, more molecules of water would occur in excited electronic states, which include apolar and paramagnetic states, generating intense bands in the spectrum. Since there is an analytical solution for the rotovibrational transitions of linear molecules, we were able to identify 16 bands relative to the excited electronic states ¹B₂ and ³A₁ in the sunspot spectrum. Density functional B3LYP/AUG-cc-pVTZ calculations of the electric and magnetic dipole are employed to discuss some consequences of the presence of excited states of water in the dynamics of sunspots and solar magnetic field.     

Water sorption in a dextran gel

The state of adsorbed water in a dextran gel has been investigated by near-infrared and gravimetric-adsorption techniques. Water-vapor adsorption (desorption) isotherms at three temperatures are reported. The calculated sorption heats are found to be markedly temperature-dependent as well as dependent on the coverage. The near-infrared spectrum (4650–9000 cm^-1) is reported, together with tentative assignments. The H₂O combination (ν + δ) band at 5184 cm_-1 has been examined as a function of relative humidity. The line-shapes of this band have been analyzed by a recently established, Fourier-inversion technique, and information on the microdynamics of the adsorbed water molecules has been resolved on the picosecond time-scale. At low and intermediate degrees of hydration, reorientational jumps take place with periods from four to six times longer than those for free water. The onset of saturation is then accompanied by the sudden removal of the reorientational jumps. A comparison of microdynamical and thermodynamic data indicates the hydration mechanism to be highly cooperative at all relative humidities.     

Vibrational analysis of peptides, polypeptides, and proteins. XVII. Normal modes of crystalline Pro-Leu-Gly-NH2, a type II beta-turn

A normal mode calculation has been done for Pro-Leu-Gly-NH2 in its crystalline type II beta-turn structure, and assignments have been made to infrared and Raman bands of this molecule and its N-deuterated derivative. Observed and calculated frequencies below 1700 cm-1 agree to within about 6 cm-1. This analysis provides a sound basis for studying the conformation dependence of the vibrational spectrum.     

A vitrational study of poly (L-proline) I and II in the far infrared.

The far infrared spectra of poly(L -proline) I (190–35 cm^?1) and II (400–35 cm^?1) were obtained in the solid state at both 300° and 110°K. A significant difference in the region below 100 cm^?1 was observed. A very intense band located at 60 cm^?1 in the infrared spectrum of form II has no counterpart in form I. This indicates the sensitivity of low-frequency vibrations to the difference in conformation assumed by both forms in the solid state. Additional bands observed in this study are correlated with ir and Raman data previously reported and tentative assignments are made using the results of normal mode calculations (in the single-chain approximation) which have been reported.     

Infrared spectra and normal vibration of β-d-glucopyranose

Infrared spectra of β-d-glucopyranose have been measured in the 4000-50 cm^-1 region and normal vibrations of β-d-glucopyranose have been investigated. The normal coordinates are treated by using the Urey-Bradley force-field, and vibrational assignments of the observed bands are made on the basis of the potential-energy distributions. The calculated vibrational-frequencies agree well with the observed frequencies in the region above 250 cm^-1.     

Laser Raman investigation of intact single muscle fibers on the state of water in muscle tissue

Laser Raman spectroscopy has been used to investigate the state of water in intact single muscle fibers of the giant barnacle (Balanus nulilus). The spectra in the region of the O-H (or O-²H) stretching modes of water in unfrozen fibers show that there is no appreciable difference between the shape and relative intensity of the Raman bands due to the water molecules located inside a muscle fiber and those of the corresponding bands in the spectrum of pure water. The presence of significant amounts of “structured” intracellular water, greater than approx. 5% of the total water content, in these fibers is thus excluded. The Raman spectra of frozen fibers have also been recorded in order to evaluate the amount of intracellular water which remains unfrozen at temperatures below the normal freezing point of water. We have been able to reproduce these spectra by assuming that the spectrum of a frozen fiber is the sum of the individual spectra of water and ice. To calculate the amount of unfrozen water from these curve fittings, it was also necessary to determine the intensities of the water and ice Raman bands relative to one another. We have found the I(ice)/I(water) ratio is 1.07 ± 0.01 for H₂O and 1.05 ± 0.03 for ²H₂O With these figures, we have calculated that for a fiber with a normal water content of 80%, 20% of the water molecules remain in the supercooled state at ?5°C, which corresponds to 1 g of water per of fiber dry weight. This amount of bound water was also found to be independent of the water content of the fibers.     

Molecular motions of polysaccharides in the solid state: dextran, pullulan and amylose.

Dextran, pullulan and amylose have been investigated by differential scanning calorimetry, thermogravimetric analysis, dynamic mechanical and dielectric spectroscopy over a wide range of temperatures and frequencies. No melting or glass transition is seen below the range of thermal degradation (about 300 degrees C) for either amylose or pullulan; only dextran shows a Tg at 223 degrees C (delta cp = 0.40 J/g deg). The viscoelastic spectrum of the 'dry' polysaccharides is characterized by a low temperature relaxation that occurs at -94, -73 and -59 degrees C, at 1 kHz, (activation energy 32, 39 and 52 kJ/mol) in dextran, pullulan and amylose respectively and is assigned to small entity local motions of the polysaccharide backbone. Absorbed water strongly modifies the relaxation spectrum, inducing a new relaxation below room temperature and dissipation regions associated with water loss above room temperature. The former appears at temperatures higher than the relaxation characteristic of the dry polymer and moves to lower temperature with increasing water content. In normal 'room humidity' conditions (about 10% absorbed water) the water-induced relaxation, attributed to the motion of complex polymer-water relaxing units, is the only observable feature in the dynamic mechanical and dielectric spectrum below room temperature.     

Fourier-transform,infrared difference-spectrometry for structural analysis of dextrans

The Fourier-transform (F.t.), infrared (i.r.) spectra of a series of branched dextrans were examined. The dextrans studied were those from the N R R L collection designated Leuconostoc mesenteroides B-1142, B-1191, B-1299 fraction S, B-1355 fraction S, B-1402, and B-1422, and Streptobacterium dextranicum B-1254 fractions S[L] and L[S]. The spectrum of a levan, N R R L L. mesenteroides B-523 fraction M, was also examined, for comparison with the spectra of the dextrans. Meaningful results were obtained by “weight-normalizing” the spectral absorbance to that of the dextran of very low degree of branching (dextran B-1254 fraction L[S]), and then subtracting this spectrum of linear dextran from each of the other polysaccharide spectra. The resulting i.r.-absorbance difference-spectra were plotted, at uniform scale-expansion across the 1800-400-cm^?1 region, resulting in difference-absorbance features at ≈ 1100 and ≈ 800 cm^?1 for all branched dextrans. These absorbance differences could be correlated to the type and degree of dextran branching, which had previously been established by permethylation analysis. It was concluded that such F.t.-i.r. difference-spectra have general application for the structural analysis of polysaccharides.     

Infrared and Raman spectroscopy of carbohydrates. Part VI: Normal coordinate analysis of V-amylose

A normal coordinate analysis of V-amylose has been performed for an isolated 6₁ helical chain. Negligible splitting from interactions of vibrations of successive residues is expected between A and E vibrational species due to the large size of the monomer unit. As a result, calculation of only the totally symmetric A modes represents an adequate approximation to the vibrational spectrum of helical polysaccharides. Using this method together with a valence force field we have obtained good agreement between the observed and calculated frequencies. In addition, the computed potential energy distribution and Cartesian displacement coordinates match previous experimental assignments, based on deuterium exchange. The analysis also supports the proposed mechanism for conversion of V-amylose to the more extended B-form. This conversion results in an observed frequency shift for the Raman line at 946 cm^?1 which is predicted by the calculations.     

Protein secondary structures in water from second-derivative amide I infrared spectra

Infrared spectra have been obtained for 12 globular proteins in aqueous solution at 20 degrees C. The proteins studied, which vary widely in the relative amounts of different secondary structures present, include myoglobin, hemoglobin, immunoglobulin G, concanavalin A, lysozyme, cytochrome c, alpha-chymotrypsin, trypsin, ribonuclease A, alcohol dehydrogenase, beta 2-microglobulin, and human class I major histocompatibility complex antigen A2. Criteria for evaluating how successfully the spectra due to liquid and gaseous water are subtracted from the observed spectrum in the amide I region were developed. Comparisons of second-derivative amide I spectra with available crystal structure data provide both qualitative and quantitative support for assignments of infrared bands to secondary structures. Band frequency assignments assigned to alpha-helix, beta-sheet, unordered, and turn structures are highly consistent among all proteins and agree closely with predictions from theory. alpha-Helix and unordered structures can each be assigned to only one band whereas multiple bands are associated with beta-sheets and turns. These findings demonstrate a method of analysis of second-derivative amide I spectra whereby the frequencies of bands due to different secondary structures can be obtained. Furthermore, the band intensities obtained provide a useful method for estimating the relative amounts of different structures.     

Complete1H NMR assignments of synthetic glycopeptides from the carbohydrate-protein linkage region of serglycins

We present complete¹H NMR assignments for two synthetic glycopeptides representative of the carbohydrateprotein linkage region of serglycin proteoglycans. The peptides are: Ser(Galp-Xylp)-Gly-Ser-Gly-Ser(Galp-Xylp)-Gly and, Ser(Galp-Xylp)-Gly-Ser(Galp-Xylp)-Gly-Ser(Galp-Xylp)-Gly. A number of 2D NMR spectra together with a 3D NOESY-TOCSY spectrum were acquired at 600 MHz to complete the assignments of the glycopeptides dissolved in water with 40% trifluoroethanol. Preliminary analysis of the NMR data suggests folded structures for the glycopeptides.A preliminary account of this work was presented at an International Symposium held at the University of Alabama at Birmingham in November, 1994 on the occasion of the 65th birthday of Professor Lennart Rodén.     

IR investigation on dehydrophenylalanine containing model peptides in helical conformation deposited on a crystal surface

Fourier transform ir spectra have been recorded for three 3₁₀‐helical and one α‐helical pentapeptides containing dehydrophenylalanine, in a thin solid film, in order to find marker bands for various secondary structures encountered in peptides containing dehydroaminoacids. The peptide solutions were deposited and dried as thin film on zinc selenide crystal surface. This convenient sampling method has provided reliable estimates of peptide secondary structure in solid state. Detailed vibrational assignments in the spectral region between 1200–1700 cm⁻¹ are reported. In this region, peptide amide I, II, and III vibrations occur. Spectra–structure correlation has been presented based on the amide modes. Comparison of the ir spectra with available crystal structure data provides qualitative support for assignments of ir bands to 3₁₀‐helical structure and α‐helical structure in dehydrophenylalanine containing pentapeptides. Band frequency assignments for 3₁₀‐helical conformation are consistent for all three peptides. All the assignments agree closely with the theoretical predictions. The spectral differences between 3₁₀‐helical peptides and the α‐helical peptide have been highlighted. These findings demonstrate that a method based on ir spectroscopy can be developed for a useful approximation of three‐dimensional structure of dehydropeptides in solid state. © 1999 John Wiley & Sons, Inc. Biopoly 50: 595–601, 1999     

Guanine residue: A normal-coordinate analysis of the vibrational spectra

The force field established for guanine is applied here to guanine-containing biopolymers by considering the model compound 9-methylguanine, in which the methyl group is taken as a dynamic unit whose mass is concentrated on the carbon. In-plane normal-mode frequencies for this model compound and its N-deuterated analog are calculated. Band frequencies observed for guanine residue in Raman biopolymer spectra, such as those for DNA, RNA, or poly(G), are associated with calculated modes having similar wavelengths. They are discussed by taking into account observed and calculated D, ¹⁵N, and ¹⁸O isotopic shifts. The atomic displacements for the normal modes corresponding to the principal bands are illustrated and a number of assignments proposed.     

Fourier transform infrared studies of ribonuclease in H2O and 2H2O solutions

Fourier transform infrared transmission spectra have been obtained of the enzyme ribonuclease in both H₂O and ²H₂O. The resolution of the spectra have been enhanced by Fourier self-deconvolution procedures. The infrared spectrum of ribonuclease changes during exchange of the enzyme's amide hydrogens for deuterium and the exchange has been followed in the amide I and amide II spectral regions. The amide I band shifts towards lower wavenumbers during both the fast and slow phases of hydrogen exchange and the interpretation of these shifts has aided the band assignments. In particular these studies have allowed an assignment to be made for the high frequency component of the β-strand absorption that differs from that proposed previously. This paper represents the first example of the use of deconvoluted Fourier transform infrared spectra in conjunction with hydrogen-deuterium exchange in order to aid in the assignment of a proteins's infrared bands.     

Far-ultraviolet optical activity of saccharide derivatives. II. Dextran,amylose, and mycodextran acetes; dextran and amylose xanthates

The ultraviolet ORD and CD spectra of amylose, dextran, and mycodextran acetates and some of thier oligomers were recorded in trifluoroethanol solution in the 300–185nm wavelength range. Similarly, the spectra of amylose and dextran xanthates in water solution were obtained in the 400–200 nm range. In the amylose acetate series, the monomer and dimer both show a normal acetyl n → π* transition in CD, while the trimer and the polymer both exhibit an additional, shorter wavelength peak. The latter is presumed to arise from a helical conformation of the amylose chain. This interpretation is substantiated by a helix–coil type transition of the CD spectra of amylose triacetate at elevated temperatures and a reversion of the anomalous CD to the normal CD upon partial deacetylation. By contrast, neither dextran acetates nor mycodextran acetate exhibit any conformational effects. The CD of dextran acetates is quite sensitive to β-1,6 and branch linkages. The ORD and CD of amylose xanthate are complex, suggesting the presence of organized structure in solution. The dextran xanthate shows only a simple ORD spectrum and no observable CD.     

Vacuum ultraviolet circular dichroism of (1 → 6)-β-d-glucan

V.u.c.d. spectra recorded for freshly prepared aqueous solutions of (1 → 6)-β)-D-glucan(pustulan) contained a single positive band near 177 nm. This band was similar in position and magnitude to the single positive band observed in the spectrum of (1 → 6)-α-D-glucan (dextran). Pustulan solutions (20 mg/ml) were observed to gel with time at 10 C. Concurrently, a negative band at 190 nm developed in the pustulan v.u.c.d. spectrum followed by a blue shift of both bands with continued aging. Crystalline films of pustulan yield spectra which resembled the blue shifted spectra of aged gels. The time dependent development of the negative band was attributed to pustulan attaining a helical conformation in solution, and the blue shift to aggregation of helices, Na⁺ and Ca²⁺ were found to accelerate gelation presumably by decreasing the activity of the aqueous solvent.     

Assignment Strategies for Large Proteins by Magic-Angle Spinning NMR: The 21-kDa Disulfide-Bond-Forming Enzyme DsbA

We present strategies for chemical shift assignments of large proteins by magic-angle spinning solid-state NMR, using the 21-kDa disulfide-bond-forming enzyme DsbA as prototype. Previous studies have demonstrated that complete de novo assignments are possible for proteins up to  ∼ 17 kDa, and partial assignments have been performed for several larger proteins. Here we show that combinations of isotopic labeling strategies, high field correlation spectroscopy, and three-dimensional (3D) and four-dimensional (4D) backbone correlation experiments yield highly confident assignments for more than 90% of backbone resonances in DsbA. Samples were prepared as nanocrystalline precipitates by a dialysis procedure, resulting in heterogeneous linewidths below 0.2 ppm. Thus, high magnetic fields, selective decoupling pulse sequences, and sparse isotopic labeling all improved spectral resolution. Assignments by amino acid type were facilitated by particular combinations of pulse sequences and isotopic labeling; for example, transferred echo double resonance experiments enhanced sensitivity for Pro and Gly residues; [2-¹³C]glycerol labeling clarified Val, Ile, and Leu assignments; in-phase anti-phase correlation spectra enabled interpretation of otherwise crowded Glx/Asx side-chain regions; and 3D NCACX experiments on [2-¹³C]glycerol samples provided unique sets of aromatic (Phe, Tyr, and Trp) correlations. Together with high-sensitivity CANCOCA 4D experiments and CANCOCX 3D experiments, unambiguous backbone walks could be performed throughout the majority of the sequence. At 189 residues, DsbA represents the largest monomeric unit for which essentially complete solid-state NMR assignments have so far been achieved. These results will facilitate studies of nanocrystalline DsbA structure and dynamics and will enable analysis of its 41-kDa covalent complex with the membrane protein DsbB, for which we demonstrate a high-resolution two-dimensional ¹³C-¹³C spectrum.     

叶冠尺度野鸭湖湿地植物群落含水量的高光谱估算模型

利用高光谱遥感技术定量估测野鸭湖湿地植被含水量,对于监测和诊断野鸭湖湿地植被的生理状况及生长趋势具有重要意义,也能够为高光谱遥感影像在野鸭湖湿地植被含水量诊断中的实际应用提供理论依据和技术支持.采用Field Spec 3野外高光谱辐射仪,获取了野鸭湖典型湿地植被冠层和叶片的光谱,并测定了对应的含水量.以上述实测数据为基础,首先以芦苇为例初步探明了不同含水量水平下典型湿地植被冠层和叶片光谱反射率的响应模式,然后采用相关性及单变量线性与非线性拟合分析技术,从冠层和叶片两种层次,对不同尺度下的含水量与“三边”参数及高光谱植被指数进行了分析拟合,并采用交叉检验中的3K-CV方法对估算模型进行了测试和检验,确立了不同尺度下野鸭湖湿地植被含水量的定量监测模型.结果表明:(1)随着含水量水平的增加,芦苇冠层与叶片光谱在可见光波段(350-760 nm)和红外波段(760-2500 nm)的反射率均呈逐渐降低趋势.(2)不同尺度含水量与选取的光谱特征参数整体上相关性较强,与“三边”参数基本上都呈极显著相关,相关系数最大达到0.906;与高光谱指数全部呈极显著相关,相关系数最小为0.455,最大达到0.919,并通过选取不同尺度上相关性最佳的光谱特征参数,分别基于“三边”参数和高光谱植被指数构建了不同尺度下的含水量估算模型.其中,冠层尺度下,黄边面积(SDy)与SRWI( Simple Ratio Water Index)的估算效果最好,估算模型分别为y=-9.462x2 -2.671x+0.608和y=0.219e1.010x;叶片尺度下,红边面积(SDr)与WI( Water Index)的估算效果最好,估算模型分别为y=0.562x+0.376和y=2.028x2 -0.476x-1.009.通过3K-CV的交叉验证,不同尺度下的含水量估算模型均取得了较为理想的预测精度,预测精度的最小值为94.92％,最大值为97.06％,表明估测模型具有较高的可靠性与普适性.(3)高光谱植被指数与含水量拟合方程的拟合度相对高于“三边”参数与之建立方程的拟合度,说明多波段组合的光谱特征参数更适合含水量的判别.     

13C NMR spectra of some naturally occurring binaphthoquinones and related compounds

¹³C NMR chemical shift assignments have been shown to be diagnostic for the establishment of the dimeric linkage of some naturally occurring binaphthoquinones. The unsymmetric ¹³C and ¹H spin-spin coupled pattern observed in the ¹H coupled ¹³C NMR spectrum of plumbagin for C-6 has also been noticed earlier with the related compound juglone. The nature of these effects has been substantiated for the first time using benzene induced solvent shifts and D₂O exchange. ¹³C chemical shift assignments of plumbagin reported earlier for C-6 and C-8 have been revised.     

Sequence-specific 1H, 13C, and 15N backbone assignment of Psb27 from Synechocystis PCC 6803

Photosystem II (PSII) is a large membrane protein complex that uses light to split water into molecular oxygen, protons, and electrons. Here we report the ¹H, ¹⁵N and ¹³C backbone chemical shift assignments for the Psb27 protein of Photosystem II from Synechocystis PCC 6803. These assignments will now provide the basis for the structural analysis of the Psb27 protein.