RELATIVE CONTRIBUTIONS OF TRYPTOPHAN and TYROSINE TO THE PHOSPHORESCENCE EMISSION OF HUMAN SERUM ALBUMIN AT LOW TEMPERATURES

Abstract— Phosphorescence emission and excitation spectra, as well as decay profiles of human serum albumin, were investigated in the wavelength regions of the tryptophan and tyrosine absorption and emission spectra in potassium phosphate buffer at 77 K. Emission and excitation spectra were found to be linear superpositions of the contributions of the tryptophan and tyrosine residues. It is suggested, therefore, that there is no significant tyrosine to tryptophan energy transfer in this protein at low temperature. The phosphorescence decay is, in general, multiexponential with lifetime components of 5.95, 2.7, and 1.2 s. The longest lifetime is characteristic of tryptophan, whereas the two short components are attributed to two types of tyrosine residues located in different environments within the protein. The latter is confirmed by a detailed analysis of the phosphorescence decay profiles determined at different emission wavelengths, and utilizing different wavelengths of excitation favoring either the tryptophan or tyrosine residues.     

TEMPERATURE DEPENDENCE OF THE PHOSPHORESCENCE LIFETIMES OF HETEROGENEOUS TRYPTOPHAN RESIDUES IN GLOBULAR PROTEINS BETWEEN 293 AND 77 K

Abstract— The phosphorescence of five globular proteins containing tryptophan residues was observed in deoxygenated neutral ethylene glycol-phosphate buffer (1:1 by volume) at 293 and 77 K. Their spectral features at 293 K are closely identical to those at 77 K apart from a lack of tyrosine phosphorescence at 293 K. and are independent of the excitation wavelength between 250 and 310 nm. From the present results. it can be concluded that the buried tryptophan residues are the only phosphorescing centers at room temperature. Their phosphorescence lifetimes were measured as a function of temperature in the range from 77 to 293 K. At room temperature, their phosphorescence lifetimes are between about 1 and 500 ms. On the basis of their temperature dependence, the heterogeneous tryptophan environments are discussed in terms of a temperature-activated nonradiative rate. We suggest that the observation of the phosphorescence characteristics of globular proteins containing tryptophan residues buried in the interior of the protein molecule at room temperature is likely to prove useful in probing the protein structure in solution.     

DSC AS A TOOL TO PREDICT EMULSION STABILITY

The crystallization of n-Hexadecane dispersed within a suitable aqueous medium has been studied by the mean of Differential Scanning Calorimetry (DSCI. The dispersion of the oil is found to be dependent on the mixing conditions: calorimetric curves exhibit one or two exothermic signals which represent different undercoolings. n-Hexadecane can be dispersed in microscopic droplets or in both microscopic and macroscopic phases. Furthermore, during an isothermal stabilization (in a particular temperature range) one can observe an oil transfer from the microscopic droplets to the macroscopic organic phase. The same phenomenon can also be induced by thermal cycling. The effect of aging was found to be in perfect agreement with the predictions that can be drawn from the calorimetric analysis of freshly made samples.     

室温下空气与金属铌和钽表面的相互作用

用XPS角分布法研究了铌和钽在室温下与干燥空气的相互作用.结果表明铌和钽表面生成了化学组成分别为Nb_2O_5和Ta_2O_5的氧化物,氧化膜内金属原子和氧原子呈混合状态,从而排除了这种相互作用仅仅是化学吸附的可能性.     

常温预辐射聚合法物理包埋葡萄糖氧化酶的研究

多数生物活性分子,如酶,抗体及药物在室温下接受辐射能后其生物活性和药力会受到不同程度的损伤。低温(-78℃)辐射对酶的辐射失活有不同程度的保护作用。为此,嘉悦憅等人曾发展了一种低温辐射包埋技术。另外,有一些酶(如过氧化氢酶、辣根过氧化物酶等)还具有辐射后失活效应,即受辐照酶样品取出辐射场后其活性仍随存放时间而不断下降。低温可以减缓后失活速率但难以完全排除。为避免低温操作的麻烦和辐射失活与后失活的损失,作者曾使用一种室温预辐射包埋技术。这一技术是将包埋过程分为两步:先使单体溶液在不断通氧下室温预辐照,然后在辐射场外与适当酶混合,通氮使其聚合,达到物理包埋的效果。     

常温预辐射聚合法物理包埋葡萄糖氧化酶的研究

Most of bioactive species such as enzyme and antibody will be damaged and lose part of their bioactivity when they are irradiated by ionizing radiation at room temperature. Low temperature could somewhat control the effect, so kaetsu and his co-workers developed an entrapping technique at low temperature ( - 78℃). On the other hand, some enzymes such as catalase and house radish peroxidase have so-called radiation-induced post-deactivation effect as well. In order to avoid the trouble in performance and loss of bioactivity, a physical ent-rapping of bioactive species via preirradiation polymerization at room temperature were deve-loped in the Authors'lab. In this work we developed this technique. The glucose oxidase was entrapped in polyacrylamide hydrogel ind uced by pre-irradiation. Experiments under several different conditions such as absorbed dose and concentrations of enzymes,monomer and crosslinking agent were carried out. The results showed that this technique can be used ex-tensively in various enzyme.     

电荷转移配合物的室温固相合成与性质研究

首次报道用室温固相反应法合成了一种有机 -多金属氧酸盐电荷转移配合物 (H2 quin) 3(PW12 O4 0 )· 4Hquin,用元素分析、红外、紫外、固体电子光谱、XRD、TG-DTA、极谱、伏安、ESR和变温磁化率等手段对其进行了研究 ,确定了该配合物的组成与结构。红外、紫外、固体电子光谱研究表明 8-羟基喹啉与杂多阴离子之间发生了电荷转移反应 ,ESR和极谱、伏安研究结果证明配合物中杂多阴离子发生了单电子还原反应 ,生成了混合价化合物。     

金属-半导体催化剂的研究Pt/SnO2上氢的室温溢流

本工作研究了Pt/SnO_2与气体分子H_2、CO之间的相互作用. 用溢流吸附反应确证室温下氢在Pt/SnO_2表面会发生一次溢流和二次溢流. 溢流氢可改变Pt/SnO_2的电导甚至能改变担体阳离子的价态.     

PHOTOCHEMICAL REACTION BETWEEN PROTEIN AND NUCLEIC ACIDS: PHOTOADDITION OF TRYPTOPHAN TO PYRIMIDINE BASES*1

Abstract— The photochemical interactions between tryptophan and nucleic acid bases were studied. When aqueous solutions of tryptophan were irradiated (Λ > 260 nm) at neutral pH in the presence of each of the nucleic acid bases, pyrimidine bases but not purine bases were altered. Air was found to decrease the rate of reaction. Two classes of photoproducts were isolated by thin layer and ion-exchange chromatography and partially characterized. One was the dihydro-pyrimidine forms of the base (see Reeve and Hopkins, 1979) and the other consisted of tryptophan-pyrimidine photoadducts. Four tryptophan-uracil and two tryptophan-thymine adducts each with a 1:1 molecular stochiometry were found. Spectroscopic measurements and a positive reaction with Ehrlich's reagent indicate that the indole nucleus remained intact, but that the pyrimidine base was reduced at the 5, 6 double bond. The absence of a positive ninhydrin reaction and the effect of pH on the quantum yield of the photoadduct formation indicated that the ionized a-amino acid group of tryptophan was involved in photoadduct formation. Indole derivatives lacking an a-amino group were also found to form photoadducts with pyrimidine bases. The experimental results are consistent with a reaction mechanism involving tryptophan excitation to the first excited singlet state as the initial event. Radical scavenging experiments indicate that tryptophan free radicals, formed by electron dissociation from the excited state, react with the ground state pyrimidine.     

EXCITED-STATE PROPERTIES OF DNA METHYLATED AT THE NV7 POSITION OF GUANINE AND ITS FREE FLUOROPHORE AT ROOM TEMPERATURE

The room-temperature optical properties of calf thymus DNA, with about 75% of its guanine residues methylated at position N-7, are compared with those of 7-methyl GMP which has the same fluorophore. The fluorescence spectrum of the methylated guanine residues depends strongly on the excitation wavelength, shifting to the blue as the wavelength increases. The fluorescence quantum yield, corrected for the contribution to absorption by the other virtually nonfluorescent residues, exhibits a pronounced drop at long excitation wavelengths relative to that for excitation at 265 nm. The degree of fluorescence polarization exhibits a weak dependence on the excitation and emission wavelengths. For 7-methyl GMP, the fluorescence spectrum is very weakly dependent on the excitation wavelength and its fluorescence quantum yield shows a moderate increase at long wavelengths. The degree of fluorescence polarization increases with increasing excitation wavelength particularly when monitoring the emission in the short wavelength region of the fluorescence spectrum. A pronounced drop of unknown origin is observed when exciting at 265 nm, which is not observed for methylated DNA. The methylated DNA data are interpreted in terms of a combination of (i) a heterogeneous environment of the methylated guanine residues, which results from sequence-dependent stacking interactions, and (ii) transfer of excitation energy from the other residues to the fluorescing methylated guanine residues. From the values of the quantum yields and those of the decay times, which we have recently reported (Georghiou et al., 1985), the following values are obtained for the radiative, k_t, and the sum of the nonradiative, σk₁, rate constants for deexcitation of the excited states of methylated DNA and its free fluorophore: 1.6 × 10⁸ s^-1 7 × 10⁷ s^-1 and 5 × 10¹⁰ s^-lvs 6 × 10⁹ s^-1. Because of energy transfer from the other residues. the k_f value for the methylated guanine residues is overestimated but their σk₁, value is not affected significantly and is by about an order of magnitude larger than that for 7-methyl GMP, apparently because of stacking interactions.     

ACTION SPECTRA OF CATION EFFECTS ON THE FLUORESCENCE POLARIZATION AND INTENSITY IN THYLAKOIDS AT ROOM TEMPERATURE

Abstract— The degree of polarization of chlorophyll- a (Chl- a ) fluorescence is known to monitor the extent of excitation migration and/or the orientation of the photosynthetic pigment molecules. We report here the effects of cations, at room temperature, on the degree of polarization of Chl- a fluorescence, and fluorescence intensity in thylakoids as a function of excitation wavelength. Observations of maxima at 650 and 675 nm in the cation-induced changes in the excitation spectrum for fluorescence at 730 and 762 nm, and, in the action spectra for the depolarization of fluorescence lead us to suggest that the regulation of the initial distribution of excitation to photosystem II involves the better coupling of Chl- b and- a in the light harvesting complex with Chl- a in the reaction center II complex.     

INTRINSIC POLARIZATION AND ROTATIONAL DEPOLARIZATION OF FLUORESCENCE OF DNA BASES IN AQUEOUS SOLUTION AT ROOM TEMPERATURE*

Abstract— A counting procedure is described for determining the polarization characteristics of very weak emission. The degree of polarization of weak emission of DNA bases in aqueous solution at room temperature is found to he high and positive, and shows an inverse correlation with their emission quantum yields which can be understood in terms of a competition between emission lifetimes and rotational relaxation times. For 5-methylcytosine the emission quantum yield can be changed by varying the acidity and the corresponding degrees of polarization show the behavior expected from Perrin's relation. The ratio τ_o/τ_rot is discussed in terms of anisotropic relaxation and calculations of τ₀.     

用凝胶色谱法在室温下测定聚对苯二甲酸丁二醇酯(PBT)分子量和分子量分布的研究

通过溶度参数等有关理论进行计算、分析及实验验证,找到了邻氯苯酚/氯仿(1/3,ml/ml)可作为聚对苯二甲酸丁二醇酯(PBT)的溶剂和室温淋洗剂。用自行制备的一系列分子量不同的PBT样品标定柱,建立了用凝胶色谱在室温下测定PBT分子量和分子量分布的方法,并得到了PBT和PS在邻氯苯酚/氯仿(1/3,ml/ml)体系中,25.0℃的[η]-M方程。发现在此GPC体系中普适标定法可以适用。     

用凝胶色谱法在室温下测定聚对苯二甲酸丁二醇酯(PBT)分子量和分子量分布的研究

 通过溶度参数等有关理论进行计算、分析及实验验证,找到了邻氯苯酚/氯仿(1/3,ml/ml)可作为聚对苯二甲酸丁二醇酯(PBT)的溶剂和室温淋洗剂。用自行制备的一系列分子量不同的PBT样品标定柱,建立了用凝胶色谱在室温下测定PBT分子量和分子量分布的方法,并得到了PBT和PS在邻氯苯酚/氯仿(1/3,ml/ml)体系中,25.0℃的[η]-M方程。发现在此GPC体系中普适标定法可以适用。     

室温交联型硅丙微胶乳的合成研究Ⅱ.改性微胶乳的粒径控制及其微观形态表征

采用种子微乳液聚合法和单体预乳化法分别合成了室温交联型有机硅改性聚丙烯酸酯微胶乳，研究了硅烷单体的添加和氧化反应对改性微胶乳粒子大小及分布的影响．结果表明：采用种子微乳液聚合法得到的硅丙微胶乳粒子大小与硅烷单体的种类．用量和添加顺序无关，平均粒径约为40-60nm．TEM照片显示出，采用种子微乳液聚合法合成的硅丙微胶乳舡子由于内部存在交联结构而导致表面形状不规则，有“乳突”现象；而采用预乳化法合成的硅丙微胶乳粒径粗大，呈规则的球形，氧化后柱径从100nm以上减小到80nm左右，粒子表面出现“绒毛”现象，这是由于硅烷组分的水解反应受到抑制而使粒子内部的交联密度降低的缘故．     

室温交联型硅丙微胶乳的合成研究II.改性微胶乳的粒径控制及其微观形态表征

采用种子微乳液聚合法和单体预乳化法分别合成了室温交联型有机硅改性聚丙烯酸酯微胶乳,研究了硅烷单体的添加和氨化反应对改性微胶乳粒子大小及分布的影响。结果表明:采用种子微乳液聚合法得到的硅丙微胶乳粒子大小与硅烷单体的种类、用量和添加顺序无关,平均粒径约为40~60nm。TEM照片显示出,采用种子微乳液聚合法合成的硅丙微胶乳粒子由于内部存在交联结构而导致表面形状不规则,有“乳突”现象;而采用预乳化法合成的硅丙微胶乳粒径粗大,呈规则的球形,氨化后粒径从100nm以上减小到80nm左右,粒子表面出现“绒毛”现象,这是由于硅烷组分的水解反应受到抑制而使粒子内部的交联密度降低的缘故。     

弱粘结煤的高温结构变化及其对合成SiC的影响

用弱粘结煤与石英砂以Acheson工艺合成了SiC ,用SEM研究了弱粘结煤高温焦炭的孔隙结构 ,并对弱粘结煤 80 0℃～ 1 60 0℃的焦炭进行了X射线衍射分析 ,计算了各温度下的微晶结构参数。研究表明 ,弱粘结煤高温焦炭孔隙率、长径比和表面积大 ,反应性好 ,可为SiC快速、大范围生长提供有利条件。微晶活化温度范围也和SiC的最佳合成温度范围基本一致。     

PHOTO-CIDNP AS A TOOL FOR THE STUDY OF THE REACTIVITY OF PHOTOSENSITIZING DRUGS: THE FUROCOUMARINS

Abstract— Several monofunctional and bifunctional furocoumarin derivatives were studied by means of the photo-CIDNP technique. Nuclear spin polarizations were observed on some of these derivatives when irradiated in acetonitrile solutions. The likely occurrence of both semi-reduced and semi-oxidized species of the FC in the reaction scheme is discussed.
When N-acetyltyrosine was added to the solution, CIDNP effects were observed on both reactants. They arose from a highly reversible pathway, as no photoproduct was polarized. On the FC part, the stronger effects were always observed on the 3,4 pyrone double bond suggesting that it must possess the highest spin density in the reaction intermediate. According to previous studies, the latter should be the FC anion or neutral radical. For the amino-acid moiety, the intensity of the polarizations was strongly dependent on the nature of the FC. This allowed the classification of the FC into three groups (zero, weak and strong intensities). As compared with their photobiological reactivity, this classification seemed to be related to the usual mono- and bifunctional distinction. This is the second example showing that photo-CIDNP could be used as a preliminary test for the study of the photosensitizing drug properties.