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1.
As a model of phospholipid bilayers in solid an oriented multilayer film (built-up film) of L-α-dipalmitoyl phosphatidylcholine (DPPC) was prepared from the monolayer by the dipping method. Structural analysis has been carried out by measuring infrared dichroism of the built-up film. The results were compared with those of the built-up film of L-α-dipalmitoyl phosphatidylethanolamine (DPPE). The tilting of the hydrocarbon chains is larger for DPPC than for DPPE. The orientation of the bisector of the two non-esterified PO bonds is closer to the film plane for DPPC than for DPPE. The strong hydrogen bonding interaction between the polar head groups was shown for DPPE, but not for DPPC. These features resemble the structural differences between dilauroyl phosphatidylethanolamine (DLPE) and dimyristoryl phosphatidylcholine (DMPC) in crystals. The hydrogen bonding interaction of DPPE found in solid remains even in the presence of water, namely, in the gel state. More closed packing of the hydrocarbon chains of solid DPPE than DPPC in solid was concluded on the basis of infrared and Raman spectra. 相似文献
2.
W L Kubasek Y Wang G A Thomas T W Patapoff K H Schoenwaelder J H Van der Sande W L Peticolas 《Biochemistry》1986,25(23):7440-7445
Raman spectra were obtained from aqueous solutions of the deoxyoligonucleotide d(CGCGAATTCGCG)2 (I), which has been suggested as a model for B-type DNA conformation. These spectra were compared with the Raman spectra of the aqueous solutions of several DNAs of natural origin taken under identical solution conditions. Since the model sequence has a high percent GC (66%), the Raman spectrum was compared with the Raman spectrum of the DNA from Micrococcus lysodeikticus (72% GC), and the spectra of the two different DNAs were found to be rather similar in both 50 mM salt and 6 M salt solutions. Computer-aided band-shape analysis of the backbone vibrational region of the Raman spectra shows the existence of several bands corresponding to different furanose ring puckers. This appears to indicate a heterogeneity of furanose ring pucker in both the model dodecamer and the native DNA. Significant differences were found in the intensity of the conformational marker band at 810 cm-1, which indicates corresponding differences in furanose ring pucker heterogeneities in these two high GC content DNAs. The Raman spectrum of the dodecamer (I) was used to analyze the Raman spectrum of the DNA inside the head of living intact salmon sperm. Sperm spectra were taken with both our conventional Raman spectrograph and a newly developed intracavity laser Raman microscope system. Although the DNA in the sperm head is required by packing considerations to be in a highly compact and condensed state, the Raman spectra of the intact sperm are almost identical with that of the model dodecamer (I) if the difference in base composition is taken into account.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
3.
Resonance Raman microspectroscopy of myeloperoxidase and cytochrome b558 in human neutrophilic granulocytes.
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N M Sijtsema C Otto G M Segers-Nolten A J Verhoeven J Greve 《Biophysical journal》1998,74(6):3250-3255
With (resonance) Raman microscospectroscopy, it is possible to investigate the chemical constitution of a very small volume (0.5 fl) in a living cell. We have measured resonance Raman spectra in the cytoplasm of living normal, myeloperoxidase (MPO)-deficient, and cytochrome b558-deficient neutrophils and in isolated specific and azurophilic granule fractions, using an excitation wavelength of 413.1 nm. Similar experiments were performed after reduction of the redox centers by the addition of sodium dithionite. The specific and azurophilic granules in both redox states appeared to have clearly distinguishable Raman spectra when exciting at a wavelength of 413.1 nm. The azurophilic granules and the cytochrome b558-deficient neutrophils showed Raman spectra similar to that of the isolated MPO. The spectra of the specific granules and the MPO-deficient neutrophils corresponded very well to published cytochrome b558 spectra. The resonance Raman spectrum of the cytoplasmic region of normal neutrophilic granulocytes could be fitted with a combination of the spectra of the specific and azurophilic granules, which shows that the Raman signal of neutrophilic granulocytes mainly originates from MPO and cytochrome b558, at an excitation wavelength of 413.1 nm. 相似文献
4.
Zhenglong Zhang Xiaorui Tian Hairong Zheng Hongxing Xu Mengtao Sun 《Plasmonics (Norwell, Mass.)》2013,8(2):523-527
We report ultrasensitive Stokes and anti-Stokes Raman spectra of 1,2-benzenedithiol monolayer on Ag film with home-made high-vacuum tip-enhanced Raman spectroscopy (HV-TERS) system. Raman peaks that were orginally very weak were observed experimentally and assigned theoretically. The local temperature was obtained based on the observed Stokes and anti-Stokes HV-TERS spectra. 相似文献
5.
A glycerol-requiring auxotroph was isolated from mutagenized Escherichia coli K-12 UFAts cells. This auxotroph was used for the specific deuteration of E. coli phospholipids. The cells were grown under high osmotic pressure (in the presence of 2.0% KCl). The membrane had a highly saturated fatty acid composition (76% phosphatidylethanolamine, 20% cardiolipin and 4% phosphatidylglycerol). The deuterium magnetic resonance spectra of coarse liposomes of the extracted phospholipids with perdeuterated glycerol incorporated into them were measured. To obtain well characterized information, phospholipid mixtures reconstituted from the deuterated and nondeuterated components at the same ratios as in the case of the total extract were used. On the analysis of the spectra, the following conclusions were drawn. (1) The whole polar region of cardiolipin is dynamically symmetric and quite rigid in the presence of phosphatidylethanolamine. (2) Although the quadrupole splittings of the deuterons at the C-2 and C-3 positions of the glycerol backbone were similar to each other, those at the C-1 position for phosphatidylethanolamine and cardiolipin are different, even in the same bilayer. (3) Furthermore, each C-1 deuteron of phosphatidylethanolamine gave rise to a doublet, suggesting the presence of two backbone conformations, between which there is slow exchange. (4) The polar head group of phosphatidylethanolamine interacts with cardiolipin and phosphatidylglycerol in different ways, which could be responsible for the different osmotic properties of the vesicles composed of them. 相似文献
6.
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
7.
大鼠大脑皮质与纹状体显微拉曼光谱的研究 总被引:1,自引:0,他引:1
用Spex-1428显微激光拉曼光谱仪测定正常大鼠大脑皮质和纹状体的激光拉曼光谱的变化,发现正常大鼠大脑皮质和纹状体的激光拉曼光谱在模式上大同小异,包含有十分丰富的生物大分子结构信息。结果如下:(1)在大脑皮质和纹状体同时出现且性状亦相似的有以下一些特征峰:808cm^-1的特征峰,对应于A型DNA的特征峰;832cm^-1和836cm^-1对就于酪氨酸(Tyr)环的振动峰;1020cm^-1和1046cm^-1相当于蛋白质中氨基酸内的C-N伸缩振动峰;1330cm^-1相当于腺嘌呤环的C=C和C-N伸缩振动峰;1544cm^-1相当于酰胺Ⅱ的N-H平面内弯曲振动和C-N伸缩援峰;1684cm^-1对应蛋白质二级结构中的转角(turn)结构。(2)大脑皮质较纹状体明显的特征峰:1092cm^-1的DNA骨架的对3称振动峰;1350cm^-1的色氨酸峰;1690cm^-1为尿嘧啶的C=O振动峰。(3)纹状体较大脑皮质明显的特征峰;1450-1463cm^-1为蛋白质CH2弯曲振动的特征峰带,纹状体在此区段有1454cm^-1峰,而大脑皮质在此区域比较低平;1490cm^-1,为鸟嘌呤的特征峰。结果显示:显微拉曼光谱揭示的大脑皮质和纹状体的生物大分子的结构成分信息十分丰富,既有共性也有差异,显微拉曼光谱是一种十分灵敏的研究手段。 相似文献
8.
High quality surface-enhanced resonance Raman (SERR) spectra were recorded from native and denatured phycocyanin and allophycocyanin on ascorbic acid treated silver hydrosols. The visible-excited SERR and resonance Raman (RR) spectra of the phycobiliproteins were very similar, indicating a predominantly electromagnetic surface enhancement mechanism. Investigation of pH-induced denaturation ofx allophycocyanin has shown that even small differences in protein/chromophore conformational are sensitively reflected by the SERR spectra. Concerning the adsorption of the protein to the metal surface, the experiments have shown that: (i) there is limited possibility for changing protein conformation during the adsorption process, (ii) there are no changes after the protein has been adsorbed onto the silver surface and (iii) for each protein an optimal activation of the silver sol has to be found for recording proper SERR spectra. The results obtained on phycobiliproteins are also discussed in connection with the interpretation of phytochrome Raman spectra. 相似文献
9.
Raman spectra of dipalmitoyl phosphatidylcholine and structurally related molecules are examined and vibrational transitions assigned for the C-C, phosphate and C-H stretching modes of these molecules. Particular emphasis is placed on the characteristics of the Raman spectra in the 2800-3000, 1000-1150 and 700-800 cm-minus 1 regions. It is found that hydrocarbon transitions dominate the spectra at the expense of those of the phosphate and choline groups. The methyl and methylene C-H stretching assignments have been clarified for the Raman spectra of phospholipid systems. 相似文献
10.
The effect of cell fixation on the discrimination of normal and leukemia cells with laser tweezers Raman spectroscopy 总被引:1,自引:0,他引:1
Laser tweezers Raman spectroscopy (LTRS) was used to characterize the effect of different chemical fixation procedures on the Raman spectra of normal and leukemia cells. Individual unfixed, paraformaldehyde-fixed, and methanol-fixed normal and transformed lymphocytes from three different cell lines were analyzed with LTRS. When compared to the spectra of unfixed cells, the fixed cell spectra show clear, reproducible changes in the intensity of specific Raman markers commonly assigned to DNA, RNA, protein, and lipid vibrations (e.g. 785, 1230, 1305, 1660 cm(-1)) in mammalian cells, many of which are important markers that have been used to discriminate between normal and cancer lymphocytes. Statistical analyses of the Raman data and classification using principal component analysis and linear discriminant analysis indicate that methanol fixation induces a greater change in the Raman spectra than paraformaldehyde. In addition, we demonstrate that the spectral changes as a result of the fixation process have an adverse effect on the accurate Raman discrimination of the normal and cancer cells. The spectral artifacts created by the use of fixatives indicate that the method of cell preparation is an important parameter to consider when applying Raman spectroscopy to characterize, image, or differentiate between different fixed cell samples to avoid potential misinterpretation of the data. 相似文献
11.
Raman and Raman optical activity (ROA) spectra were collected for four RNA oligonucleotides based on the EMCV IRES Domain I to assess the contributions of helix, GNRA tetraloop, U·C mismatch base pair and pyrimidine-rich bulge structures to each. Both Raman and ROA spectra show overall similarities for all oligonucleotides, reflecting the presence of the same base paired helical regions and GNRA tetraloop in each. Specific bands are sensitive to the effect of the mismatch and asymmetric bulge on the structure of the RNA. Raman band changes are observed that reflect the structural contexts of adenine residues, disruption of A-form helical structure, and incorporation of pyrimidine bases in non-helical regions. The ROA spectra are also sensitive to conformational mobility of ribose sugars, and verify a decrease in A-type helix content upon introduction of the pyrimidine-rich bulge. Several Raman and ROA bands also clearly show cooperative effects between the mismatch and pyrimidine-rich bulge motifs on the structure of the RNA. The complementary nature of Raman and ROA spectra provides detailed and highly sensitive information about the local environments of bases, and secondary and tertiary structures, and has the potential to yield spectral signatures for a wide range of RNA structural motifs. 相似文献
12.
The major lipids of Tetrahymena membranes have been purified by thin-layer and high pressure liquid chromatography and the phosphatidylethanolamine and aminoethylphosphonate lipids were examined in detail. 31P-NMR, X-ray diffraction and freeze-fracture electron microscopy were employed to describe the phase behavior of these lipids. The phosphatidylethanolamine was found to form a hexagonal phase above 10°C. The aminoethylphosphonate formed a lamellar phase up to 20°C, but converted to a hexagonal phase structure at 40°C. Small amounts of phosphatidylcholine stabilized the lamellar phase for the aminoethylphosphonate. 31P-NMR spectra of the intact ciliary membranes were consistent with a phospholipid bilayer at 30°C, suggesting that phosphatidylcholine in the membrane stabilized the lamellar form, even though most of the lipid of that membrane prefers a hexagonal phase in pure form at 30°C. 31P-NMR spectra also showed a distinctive difference in the chemical shift tensor of the aminoethylphosphonolipid, when compared to that of phosphatidylethanolamine, due to the difference in chemical structure of the polar headgroups of the two lipids. 相似文献
13.
14.
We have constructed a Raman microscope that has enabled us to obtain resonance Raman vibrational spectra from single photoreceptor cells. The laser beam which excites the Raman scattering is focused on the outer segment of the photoreceptor through the epiillumination system of a light microscope. Raman scattering from the visual pigment in the photoreceptor is collected by the objective and then dispersed onto a multichannel detector. High-quality spectra are recorded easily from individual outer segments that are 5 x 50 micrometer in size, and we have obtained spectra from cells as small as 1 x 10 micrometer. We have used the Raman microscope to study photostationary steady-state mixtures in pigments from toad (Bufo marinus) and goldfish (Carassius auratus) photoreceptors; these photoreceptors were frozen in glycerol glasses at 77 degrees K. Comparison of our toad red rod spectra with previously published spectra of bovine rod pigments demonstrates that the conformation of the chromophore in the first photointermediate, bathorhodopsin, is sensitive to variations in protein structure. We have also studied the first photointermediate in the goldfish rod photostationary steady-state. This bathoporphyropsin has a much lower ethylenic stretching frequency (1,507 cm-1) than that observed in the toad and bovine bathoproducts (approximately 1,535 cm-1). Preliminary results of our work on goldfish cone pigments are also reported. These are the first vibrational studies on the vertebrate photoreceptors responsible for color vision. 相似文献
15.
In situ mapping of nitrifiers and anammox bacteria in microbial aggregates by means of confocal resonance Raman microscopy 总被引:1,自引:0,他引:1
Pätzold R Keuntje M Theophile K Müller J Mielcarek E Ngezahayo A Anders-von Ahlften A 《Journal of microbiological methods》2008,72(3):241-248
In the present work the exploration of microbial communities by confocal resonance Raman microscopy (CRRM) is reported. Using the resonance Raman effect of cytochrome c (Cyt c) we were able to record the microbial distribution of nitrifiers and anammox bacteria directly in their natural environment without the need of sample preparation. For this new non-invasive investigation a reference database of bacteria assumed to be found in microbial aggregates obtained from biological wastewater treatment was created. Reference spectra of enriched cultures of the interesting bacteria were taken by means of optical tweezers. Significant spectra were achieved in less than 1 s (excitation wavelength 532 nm, laser power 9 mW) due to the resonance Raman effect. In addition, the impact of different parameters on the reference spectra, such as integration time and culture conditions, was analysed. We successfully demonstrate the grouping of bacteria down to strain level based on the heterogeneity of the resonance Raman spectra of the heme protein Cyt c. 相似文献
16.
17.
To explore the biochemical differences between brain cancer cells Astrocytoma and normal cells Astrocyte, we investigated
the Raman spectra of single cell from these two cell types and analyzed the difference in spectra and intensity. Raman spectrum
shows the banding pattern of different compounds as detected by the laser. Raman intensity measures the intensity of these
individual bands.
The Raman spectra of brain cancer cells was similar to those of normal cells, but the Raman intensity of cancer cells was
much higher than that of normal cells.
The Raman spectra of brain cancer Astrocytoma shows that the structural changes of cancer cells happen so that many biological
functions of these cells are lost. The results indicate that Raman spectra can offer the experimental basis for the cancer
diagnosis and treatment. 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.
E. Mayer D.J. Gardiner R.E. Hester 《Biochimica et Biophysica Acta (BBA)/General Subjects》1973,297(2):568-570
The Raman spectra of cyanocobalamin (vitamin B12) and dicyanocobinamide have been obtained from aqueous solutions at concentrations of approx. 1 10−4 M. The spectra were excited by laser radiation coincident in wavelength with the visible absorption, this resulting in selective enhancement of some vibrational modes through the rigorous resonance Raman effect. In spite of substantial chemical differences, cyanocobalamin and dicyanocobinamide give essentially identical spectra, indicating that only those modes associated with the common corrin ring system are resonance enhanced. 相似文献
20.
2,2,5,5-Tetramethyl-3-pyrrolin-1-yloxy-3-carboxamide (tempyo) labeled bovine serum albumin and cytochrome c at different pH values were prepared and investigated using Raman-resonance Raman (RR) spectroscopy and surface enhanced Raman scattering (SERS) spectroscopy. The Raman spectra of tempyo labeled proteins in the pH 6.7-11 range were compared to those of the corresponding free species. The SERS spectra were interpreted in terms of the structural changes of the tempyo labeled proteins adsorbed on the silver colloidal surface. The tempyo spin label was found to be inactive in the Raman-RR and SERS spectra of the proteins. The alpha-helix conformation was concluded to be more favorable as the SERS binding site of bovine serum albumin. In the cytochrome c the enhancement of the bands assigned to the porphyrin macrocycle stretching mode allowed the supposition of the N-adsorption onto the colloidal surface. 相似文献