The conformation alteration of mouse hepatic histones after reacting with nicotine in vitro

UV differential spectroscopy, fluorescence spectroscopy and circular dichroism (CD) spectroscopy assays have been applied to studying the conformation alteration of mouse hepatic histones H1 and H3 after reacting with nicotine in vitro. The results indicate that their conformation changes from regular form to random form with the increasing reaction dose of nicotine. The adduction of nicotine or its metabolites with histones H1 and H3 accounts for the conformation alteration. Nicotine may affect the structure, function and expression of genes of chromosome by changing the conformation of histones.     

十二烷基硫酸钠在水滑石层间的插层行为研究

采用¹³C和²⁷Al固体核磁共振表征方法结合XRD及热分析等表征技术,对十二烷基硫酸钠（NaDS）柱撑的水滑石（Mg₂Al-NO^-3）的分子结构进行了研究。结果表明,柱撑物中的有机相存在两种不同的构形,外表面吸附的DS^-主要是以活动性较好的无序构形状态存在,进入层间的DS^-主要是以活动性差的有序构形状态存在。NaDS柱撑LDHs中有序构形的量随DS^-溶液浓度的增加而增加,当溶液中DS^-的物质的量达到水滑石阴离子交换容量（AEC）的2倍或更大时,层间DS^-的量保持为57%左右。NaDS柱撑水滑石的铝核周围的配位为单一的六配位环境,此环境在柱撑前后没有发生明显变化。     

Study the effects of divalent metallic ions on the combination of DNA and histones with fluorescence anisotropy assays

The effects of divalent ions(Mn2 ,Mg2 ,Ca2 ) on the interaction between DNA and histone are studied using a fluorescence anisotropy assay. Fluorescence anisotropies of DNA and DNA-histone in the presence of divalent ions(Mn2 ,Mg2 ,Ca2 ) are measured. The results indicate that histone reduces the fluorescence anisotropy of lambda DNA while the divalent ions(Mn2 ,Mg2 ,Ca2 ) significantly enhance the fluorescence anisotropy. Compared to the case of DNA incubated with histone alone,there are more histones binding to DNA when divalent ion,histone and DNA are incubated together. We also find that Mn2 makes the DNA-histone complexes more condensed than the other ions do.     

Binding of vanadium compounds perturbs conformation and aggregation state of insulin

The interactions between zinc-free insulin and vanadium compounds, NaVO3, VO(acac)2 and VO(ma)2, have been investigated by fluorescence spectroscopy, circular dichroism (CD) and Fourier-transformed infrared (FT-IR) spectroscopy. The results showed that binding of vanadium compounds produced a static quenching of the intrinsic fluorescence of insulin. The apparent association constants were determined to be (0.17±0.01)×104 L*mol-1 for NaVO3, (2.8±0.2)×104 L*mol-1 for VO(acac)2, and (4.0±0.1)×104 L*mol-1 for VO(ma)2, respectively. The light scattering intensity of insulin decreased upon incubation with the vanadium compounds, suggesting the disaggregation of insulin. The attenuation of the band at 273 nm of insulin CD spectra also supported the disaggregation of insulin observed above. A new band at 1650～1653 cm-1 appeared in the FT-IR spectra of insulin upon incubation with the vanadium compounds, indicating the formation of an α-helix structure at B (9-19) motif. This α-helix structure suggests a structural change of insulin from an extended conformation (T state) to a helical conformation (R state), which is essential for binding of insulin to its receptor. In conclusion, binding of vanadium compounds results in conformational changes and disaggregation of insulin. These changes might account for the enhancement of binding affinity for insulin to its receptor in the presence of vanadium compounds.     

Thermodynamic study on conformation change of carbonic anhydrase

The effect of pHs on structural stability of the zinc-containing metalloenzyme carbonic anhydrase has been studied by means of differential scanning calorimetry. The rule of the conformational change for the enzyme with pH change has been found by thermodynamic analysis. The experimental results also provide valuable information: in the pH range from 5.26 to 9.04, two independent endothermal peaks were observed on DSC thermogram. It shows the existence of two structural domains in the molecule. The transition temperature and enthalpy of the two domains are different.     

疏水缔合聚合物溶液形态的研究

用环境扫描电镜(ESEM)和动态光散射仪(DLS)对疏水缔合聚合物溶液形态进行了分析。结果表明,随着NaCl浓度的增加,实验所合成的疏水缔合聚合物的表观粘度先降低后增加而后再降低,但仍保持很高粘度。疏水缔合聚合物在去离子水体系中无论浓度高低均形成了空间网状结构。盐水体系中,当NaCl浓度为1000mg/L时所形成的结构与去离子水中的结构类似,均为网状结构,而在NaCl浓度为5000mg/L时,ESEM照片显示疏水缔合聚合物的结构为致密的树枝状结构。疏水缔合聚合物的浓度越大,其表观流体力学半径就越大,从而造成其增粘能力越强。     

Study the effects of metallic ions on the combination of DNA and histones with molecular combing technique

The effects of monovalent (Na^ , K^ ) and divalent (Mg^2 , Ca^2 , Mn^2 ) ions on the interaction between DNA and histone are studied using the molecular combing technique. λ-DNA molecules and DNA-histone complexes incubated with metal cations (Na^ , K^ , Mg^2 , Ca^2 , Mn^2 ) are stretched on hydrophobic surfaces, and directly observed by fluorescence microscopy. The results indicate that when these cations are added into the DNA solution, the fluorescence intensities of the stained DNA are reduced differently. The monovalent cations (Na^ , K^ ) inhibit binding of histone to DNA. The divalent cations (Mg^2 , Ca^2 , Mn^2 ) enhance significantly the binding of histone to DNA and the binding of the DNA-histone complex to the hydrophobic surface. Mn^2 also induces condensation and aggregation of the DNA- histone complex.     

Study of the interaction of DNA and histones by spin-stretching and droplet evaporation

Molecular combing is a powerful and simple method for aligning DNA molecules onto a surface. Using this technique combined with fluorescence microscopy, DNA-histone complexes are stretched on a hydrophobic polymethyl methacrylate (PMMA) surface and observed directly. We have developed a new method to stretch single DNA-histone complexes, termed spin-stretching. The results show that the histones markedly enhance DNA binding to the PMMA surface. DNA winds around the histones and therefore decreases in length. The number of histones that bind to each DNA molecule is found to correlate with the histone concentration. The combed DNA-histone complexes are found to depend on two factors: the binding force on the surface and the centrifugal force at its local position. Na+ ions should compete with histones for binding to DNA; however, the observed competitive binding effect of Na+ ions at low concentrations was negligible.     

Monitoring the binding of metal cations and histones to DNA in real time using fluorescence assays

The binding of cations (Na +,K +,Mg 2+,Ca 2+,Mn 2+) and histones to DNA can be studied using fluorescence assays.Here,we measured the fluorescence intensity and fluorescence anisotropy of DNA and DNA-histone complexes in the presence of cations.We demonstrate that when different cations are added into a DNA solution,the fluorescence intensities of the stained DNA are reduced by different amounts.Compared with divalent cations,monovalent cations had a weaker effect on fluorescence intensity and fluorescence anisotropy.Divalent (Mn 2+,Mg 2+,Ca 2+) cations markedly enhanced the fluorescence anisotropy of DNA.The binding modes of monovalent and divalent cations to DNA may be different.Divalent cations can change the structure of DNA molecules,or promote the assembly of DNA strands.The addition of histones causes DNA condensation,which mostly occurs during the first few seconds.Cation binding to DNA is abrupt,and is much faster than that of histones.     

双金属Hg2+和Cu2+对木瓜蛋白酶活性与构象的影响

研究双金属Hg2+和Cu2+对木瓜蛋白酶活性与构象的影响.利用FT-IR、荧光发射以及紫外吸收光谱探讨Hg2+和Cu2+处理与木瓜蛋白酶二级结构变化的关系.研究结果表明:金属离子与木瓜蛋白酶活性之间存在剂量-效应关系,表现出低剂量促进,高剂量抑制的Hormesis现象.低浓度下,双金属Hg2+和Cu2+表现出协同激活效应;高浓度下,Cu2+的添加屏蔽了Hg2+的抑制作用.双金属离子浓度为10-6 mol/L Hg2+和10-8 mol/L Cu2+时,对酶的激活效应最大,其处理的木瓜蛋白酶的有序结构(α-螺旋和β-折叠)含量最高,二级结构最稳定,酶与底物亲和力最强,活性最高.当双金属离子浓度为10-4 mol/L Hg2+和10-4 mol/L Cu2+时,其抑制力最强,处理的木瓜蛋白酶有序结构含量最低,二级结构破坏,活性最低.木瓜蛋白酶分子构象的有序度与其活性呈正相关.     

大腹园蛛丝腺蛋白二级结构的圆二色性研究

用圆二色光谱研究了大腹园蛛各丝腺蛋白的分子构象、大、小囊状腺体蛋白的分子构象以无规卷曲和β-折替结构为主;而其管状腺、集合状腺、鞭毛状腺蛋白的 CD 谱均显示出有α-螺旋结构的特征。用计算法所得这五种丝腺体蛋白的二级结构中含有6.2%～13.9%不等的β-转角,这对蛛丝蛋白形成独特的结构起着重要作用。还研究了放置时间对这五种丝腺体蛋白构象的影响。     

pH值对鲑鱼降钙素溶液构象的影响

采用傅里叶变换红外光谱和圆二色谱,测定鲑鱼降钙素在不同pH值水溶液中的构象,并对各种二级结构的含量进行计算.结果表明pH值对鲑鱼降钙素溶液构象有较大影响,pH值从2.0升高到8.0时鲑鱼降钙素溶液中存在β-折叠、β-转角和无序结构,无α-螺旋结构,且各种二级结构的含量随pH值增加而发生变化.当pH值增加到9.0时鲑鱼降钙素溶液中出现α-螺旋结构,且β-折叠和无序结构含量降低到最小值,β-转角含量增加到最大值.以上研究结果提示较低的pH值有助于鲑鱼降钙素的稳定,为进一步阐明pH值对鲑鱼降钙素稳定性的影响提供了依据.     

变性剂和缓冲系统对适冷蛋白酶MCP-01和中温蛋白酶BP-01构象影响的圆二色光谱分析何海伦， 陈秀兰*

运用圆二色（CD）光谱分析了适冷蛋白酶MCP-01和中温蛋白酶BP-的二级结构。与BP-01相比,MCP-01具有较低的α螺旋含量和较高的无规则卷曲,表明适冷蛋白酶MCP 01的结构可能具有较高的柔性和较低的稳定性。测定MCP-01和BP-01的变性温度表明,MCP-01的变性温度比BP 01 低10.7℃。测定了变性剂盐酸胍（GuHCl）、2,2,2 三氟乙醇（TFE）对MCP-01和BP-01结构的影响。盐酸胍使MCP-01和BP 01变构的浓度分别为1.0mol/L和3.0mol/L,TFE造成MCP 01和BP 01开始变构的含量分别为20%和30%。这表明,与中温酶相比,变性剂在较低浓度下就可使适冷酶变构,从而造成活性的丧失。检测了两种蛋白酶在保温过程中构象的变化情况。不同缓冲系统明显影响MCP 01的构象稳定性,MCP 01在不同缓冲液中构象稳定性为：碳酸缓冲液相似文献   

不同pH值的酸处理溶菌酶溶液的拉曼光谱分析

分析了pH 5.0到pH 1.0的溶菌酶溶液的激光拉曼谱,酰胺Ⅰ振动谱带分别出现在1660、1656、1657、1660和1655 cm^-1,酰胺Ⅲ振动谱带分别出现在1255、1257、1263、1261、1260 cm^-1和1301、1302、1302、1302、1301 cm^-1,并计算了它们相对强度的变化.由此定性可知,该蛋白质分子的肽链主要由α-螺旋和无规则卷曲组成.在酸化的过程中,溶菌酶的构象基本一直保持稳定,但当pH＜2.0时,该溶菌酶开始变性.在pH=5.0时酪氨酸为“暴露式”,当pH=4.0-2.0时为“埋藏式”,而到了pH=1.0时酪氨酸的双峰已基本消失了.色氨酸的1361 cm^-1的强度随酸性的增强而减弱,由“埋藏式”逐渐转化为“暴露式”.硫-硫桥键的振动谱带按pH5.0到pH1.0的顺序分别在508、508、507、507和509cm^-1,且强度基本不变,表明在酸环境中,硫-硫键部位的几何构型都是扭曲-扭曲-扭曲,pH值的改变对几何构型无影响.     

Early association of electrocardiogram alteration with infarct size and cardiac function after myocardial infarction

OBJECTIVE: Myocardial infarction (MI) is the main cause of heart failure, but the relationship between the extent of MI and cardiac function has not been clearly determined. The present study was undertaken to investigate early changes in the electrocardiogram associated with infarct size and cardiac function after MI. METHODS: MI was induced by ligating the left anterior descending coronary artery in rats. Electrocardiograms, echocardiographs and hemodynamic parameters were assessed and myocardial infarct size was measured from mid-transverse sections stained with Masson's trichrome. RESULTS: The sum of pathological Q wave amplitudes was strongly correlated with myocardial infarct size (r = 0.920, P < 0.0001), left ventricular ejection fraction (r = -0.868, P < 0.0001) and left ventricular end diastolic pressure (r = 0.835, P < 0.0004). Furthermore, there was close relationship between MI size and cardiac function as assessed by left ventricular ejection fraction (r = -0.913, P < 0.0001) and left ventricular end diastolic pressure (r = 0.893, P < 0.0001). CONCLUSION: The sum of pathological Q wave amplitudes after MI can be used to estimate the extent of MI as well as cardiac function.