天花粉蛋白小结构域N端同源片段的溶液构象,T18肽的圆二色性…

应用圆二色性,内源荧光和疏水荧光探针法进一步研究Ｔ１８肽的溶液构象及其相互转化,发现Ｔ１８肽在水溶液中为β折叠结构,且在高浓度（〉１ｍｇ／ｍｌ）时形成疏水聚合物,Ｌｙｓ１５和Ｉｌｅ３－Ｉｌｅ４是形成β折叠疏水簇的关键因素,并讨论了蛋白质链和溶剂环境对肽段二级结构的调制作用。     

NMR法研究天花粉蛋白TDK肽片段的溶液构象

用ＨＮＭＲ法测定ＴＤＫ肽在Ｈ２Ｏ（ＨＯＤＫ）,５０％六氟丙醇（ＦＰＤＫ）和２ｍｏｌ／ＬＧｕ．ＨＣｌ（ＧＵＤＫ）溶液构象。在ＨＯＤＫ和ＦＰＤＫ中,ＴＤＫ肽的两段序列Ａｓｐ０－Ｉｌｅ４,Ｓｅｒ９－Ｉｌｉ１７分别具有较稳定的α－螺旋含量;而ＧＵＤＫ的ＳＡＬＳ序列仍能检测到有序残存结构。并假设ＳＡＬＳ序列是肽链形成二级结构的原始核心。     

NMR法研天花粉蛋白T14肽片段的溶液构象

本文报道用ＨＮＭＲ法测定Ｔ１４肽在ＤＭＳＯ（ＭＳ１４）,Ｈ２Ｏ（ＨＯ１４）,５０％甲醇（ＭＥ１４）和５０％六氟异丙醇（ＦＰ１４）中的溶液构象。通过ＮＯＥ效应,偶合常数,Ｈ／Ｄ交换速率表征和定位二级结构和疏水域,计算两面角φ和螺旋形成几率。结果表明序列Ｓｅｒ９－Ｉｌｅ１７在ＨＯ１４中较为有序,存在强疏水作用,推测是类似螺旋样的亚稳结构;而在ＤＭＳＯ中序列ＳＡＬＳＫＱ存在形成α－螺旋结构的可能性,且六     

天花粉蛋白小结构域N端同源片段的溶液构象：T18肽的圆二色性和荧光研究

应用圆二色性,内源荧光和疏水荧光探针法进一步研究Ｔｌ８肽的溶液构象及其相互转化。发现Ｔ１８肽在水溶液中为β折叠结构,且在高浓度（＞１ｍｇ／ｍｌ）时形成疏水聚合物;Ｌｙｓ１５和Ｉｌｅ３－Ｉｌｅ４是形成β折叠疏水簇的关键因素。并讨论了蛋白质肽链和溶剂环境对肽段二级结构的调制作用。     

在不同物理化学条件下天花粉蛋白的圆二色性研究

用圆二色谱研究了天花粉蛋白在酸性和碱性溶液中的构象,并观察了在十二烷基磺酸钠(SDS)溶液中天花粉蛋白分子构象的变化,用Chen,Yang与Martinez等人的方法计算了各种条件下天花粉蛋白二级结构含量.结果表明:在很宽广的pH范围内(从pH2到pH12)分子构象基本上是稳定的.但是,在强碱性条件下即pH>12(室温)或pH11.5(40℃)螺旋含量显著减少.在碱性条件下,于250nm附近出现一新的CD带,它的强度与离解的酪氨酸残基个数呈线性关系,说明这一新带主要是由离解的酪氨酸残基产生的.     

天花粉蛋白T18肽片段的溶液构象及T14,TDK和T18肽…

用＾１Ｈ ＮＭＲ法测定Ｔ１８肽在ＤＭＳＯ（ＭＳ１８）和５０％六氟异丙醇（ＦＰ１８）中的溶液构象。ＭＳ１８含有Ｉｌｅ３￣Ｇｌｕ７和Ａｌａ１２￣Ｇｌｎ１６两段β－折叠链;而ＦＰ１８则转变为α－螺旋结构。综合分析Ｔ１４,Ｔ１８和ＴＤＫ三个模型肽的结构性质和稳定性,比较肽链序列和溶剂作用,提出肽链局部优势结构的概念,并据此讨论天花粉蛋白小结构域折叠起始过程。     

HMR法研究天花粉蛋白TDK肽片段的溶液构象

用１ＨＮＭＲ法测定ＴＤＫ肽在Ｈ２Ｏ（ＨＯＤＫ）,５０％六氟丙醇（ＦＰＤＫ）和２ｍｏｌ／ＬＧｕ·ＨＣｌ（ＧＵＤＫ）中的溶液构象。在ＨＯＤＫ和ＦＰＤＫ中,ＴＤＫ肽的两段序列Ａｓｐ０～Ｉｌｅ４,Ｓｅｒ９～Ｉｌｅ１７分别具有较稳定的α－螺旋含量;而ＧＵＤＫ的ＳＡＬＳ序列仍能检测到有序残存结构。并假设ＳＡＬＳ序列是肽链形成二级结构的原始核心。     

天花粉蛋白的作用机制—RNA N—糖苷酶型

核糖体失活蛋白(ribosome-inactivating protein,RIP)是一类广泛分布于植物界的能抑制真核细胞核糖体功能的毒蛋白。其作用的分子机制有两类:(1)RNA水解酶型,如帚曲霉素(α-sarcin),专一水解28SrRNA第4325—4326位之间的磷酸二酯键;(2)RNA N-糖苷酶(RNA N-glycosidase)型,如蓖麻蛋白A     

天花粉蛋白T18肽片段的溶液构象及T14、TDK和T18肽构象比较

用１ＨＮＭＲ法测定Ｔ１８肽在ＤＭＳＯ（ＭＳ１８）和５０％六氟异丙醇（ＦＰ１８）中的溶液构象．ＭＳ１８含有Ｉｌｅ３～Ｇｌｎ７和Ａｌａ１２～Ｇｌｎ１６两段β－折叠链;而ＦＰ１８则转变为α－螺旋结构。综合分析Ｔ１４,Ｔ１８和ＴＤＫ三个模型肽的结构性质和稳定性,比较肽链序列和溶剂作用,提出肽链局部优势结构的概念,并据此讨论天花粉蛋白小结构域折叠起始过程．     

Toxicity of Pyroglutaminated Amyloid β-Peptides 3(pE)-40 and -42 Is Similar to That of Aβ1-40 and -42

An N-terminal truncated isoform of the amyloid beta-peptide (A beta) that begins with a pyroglutamate (pE) residue at position 3 [A beta3(pE)-42] is the predominant isoform found in senile plaques. Based upon previous in vitro studies regarding A beta N-terminal truncated isoforms, it has been hypothesized that A beta3(pE)-x isoforms may aggregate more rapidly and become more toxic than corresponding Abeta1-x peptides. However, the toxicity and aggregation properties of A beta3(pE)-42 and A beta3(pE)-40 have not previously been examined. After initial solubilization and 1-week preaggregation of each peptide at 37 degrees C and pH 7.4, the toxicity of 5-50 microM A beta3(pE)-42 was similar to that of A beta1-42. Moreover, the toxicity of A beta3(pE)-40 paralleled that induced by A beta1-40 in both 1 day in vitro (DIV) cortical and 7 DIV hippocampal cells. Circular dichroism spectra did not reveal major differences in secondary structure between aged A beta1-42, A beta3(pE)-42, A beta3(pE)-40, and A beta1-40 or freshly solubilized forms of these peptides. Overall, the data indicate that the loss of the two N-terminal amino acids and the cyclization of glutamate at position 3 do not alter the extracellular toxicity of A beta.     

Circular dichroism and absorption studies of (-)-2,2'-dimethyl-4,5-(1-naphthyl)-1,3-dioxolane in terms of vibronic coupling theory

The absorption and circular dichroism (CD) spectra of (-)-2,2'-dimethyl-4,5-(1-naphthyl)-1,3-dioxolane (DND) were studied in the energy region 30,000 cm(-1) to 50,000 cm(-1). The DND ketal is treated as a naphthalene dimer and its spectra are interpreted in terms of a vibronic dimer model which includes the (1)L(a) and (1)B(b) states of the naphthalene chromophore. To fix the most stable conformation of DND molecule, the MNDO/AM1, RHF/6-31G, and SVWN5, BPW91 methods are employed with 6-31G and 6-31G(d',p') basis sets. All the methods are shown to yield the DND geometry that is entirely consistent with the CD and absorption spectra studied.     

金属硫蛋白及其结构域突变体的紫外和圆二色光谱学研究

采用 p GEX- 4T- 1融合表达载体高效表达所得的金属硫蛋白及其结构域突变体 ,包括 α结构域 ,β结构域 ,α- KKS- α和 β- KKS- β( KKS为金属硫蛋白结构域之间的天然连接区氨基酸 ) ,经纯化和纯度鉴定后 ,利用紫外和圆二色光谱进行结构研究 .在脱金属的上述蛋白中 ,固定 p H为中性 ,改变加入 Cd2 + 的比例 ,或固定 Cd2 + 浓度 ,逐渐调节 p H至中性 ,观察紫外和圆二色光谱中镉硫金属簇吸收峰的形成 .研究结果表明 :镉硫金属簇的形成依赖于加入金属的比例和 p H值 ,所有蛋白均于p H3.1 5以上开始形成明显的吸收峰 .紫外图谱中的吸收峰位于 2 54nm附近 ,但在圆二色图谱中不同蛋白形成的峰的位置不同 ,MT,α结构域和 α- KKS- α在 2 2 5nm和 2 58nm处有吸收 ,β结构域在 2 60 nm处有吸收 ,而 β- KKS- β在 2 4 5nm处有吸收 ;向 α结构域 ,β结构域 ,MT,α- KKS- α和 β-KKS- β中分别加入 4eq,3eq,7eq,8eq和 6eq( eq:equivalent,当量 )的 Cd2 +时 ,吸收峰可达到最大值 .同时发现 α结构域的吸收峰强于 β结构域 ,而且双结构域突变体的镉硫金属簇则明显强于相应的单结构域突变体 ,这表明吸收峰的强弱与金属结合力的大小相关 ,而且结构域之间存在相互作用 ,从而影响与金属的结合 .     

Conformational study of the preferred conformations of the peptide sequence VP3(110-121) of HAV by circular dichroism and molecular mechanics

Summary A conformational analysis of the fragment 110–121 of VP3 coating protein of the hepatitis A virus was carried out using circular dichroism spectroscopy and computational studies. The latter studies indicate the tendency of the peptide to adopt hairpin-type structures. Circular dichroism experiments indicate that, in spite of the fact that the isolated peptide exhibits no structure under different experimental conditions, negatively charged liposomes induce a secondary structure that agrees with the results of the computational study.     

Conformational study of the preferred conformations of the peptide sequence VP3(110-121) of HAV by circular dichroism and molecular mechanics

A conformational analysis of the fragment 110–121 of VP3 coating protein of the hepatitis A virus was carried out using circular dichroism spectroscopy and computational studies. The latter studies indicate the tendency of the peptide to adopt hairpin-type structures. Circular dichroism experiments indicate that, in spite of the fact that the isolated peptide exhibits no structure under different experimental conditions, negatively charged liposomes induce a secondary structure that agrees with the results of the computational study.     

生长调节物质对栝楼毛状根生长和天花粉蛋白合成的影响

添加GA3和CCC虽然影响栝楼毛状根的生物量积累 ,但是有利于促进天花粉蛋白 (TCN)的合成 ,当GA3的添加量为 2mg/L时 ,天花粉蛋白含量增加了 18 9% ,添加 1mg/L～ 2mg/LCCC的天花粉蛋白提高了 2 8%。分别添加维生素B1或B5会延长毛状根的生长迟滞期 ,但是 ,后期的生长非常迅速 ,并可以促进天花粉蛋白的合成。同时添加维生素B1和B5可以缩短生长迟滞期 ,并能促进毛状根的生长 ,同时添加1mg/LB1 4mg/LB5有利于促进天花粉蛋白总量的增加。     

Conformational change of poly( -lysine) induced by lipid vesicles of dilauroylphosphatidic acid

The effect of negatively charged dilauroylphosphatidic acid (DLPA) vesicles on the conformation of poly( -lysine) was investigated by circular dichroism measurements. DLPA vesicles induced a confomiational change Of poly( -lysine) from the random coil to β-structure in 5 mM Tes, pH 7.0. The fraction of induced β-structure (F_β) was determined via a procedure of curve fit the observed spectra to the reference spectra. F_β increased linearly with the molar ratio, r, of DLPA to lysine residues up to r 0.7, and reached a saturation value of 1 at r > 1. Within the range 0.7 r 1, precipitation occurred. The effect of dilution of the negative charge on vesicle membranes was examined by mixing DLPA with dilauroylphosphatidylcholine (DLPC). Although the β-structure Of poly -lysine) was also induced by mixed vesicles, the saturation value of F_β decreased with decreasing DLPA content in mixed vesicles. The variation in saturation value of F_β with the composition of mixed vesicles was interpreted in terms of the change in average distance between DLPA head groups in mixed vesicles.     

Studies on the interaction of copper complexes of (-)-epicatechin gallate and (-)-epigallocatechin gallate with calf thymus DNA

The interaction of copper complexes of (−)-epicatechin gallate (ECG) and (−)-epigallocatechin gallate (EGCG) with calf thymus DNA (ct-DNA) was investigated by UV-visible (UV-Vis), fluorescence and circular dichroism along with melting studies. It was observed that both copper complexes quench the fluorescence intensity of ethidium bromide bound ct-DNA upon binding, resulting in a ground state complex formation by a static quenching process. The binding constants evaluated from fluorescence data were supported by the UV-Vis study. The values ranged from 0.84 to 1.07 × 10⁵ M⁻¹ and 1.14 to 1.04 × 10⁵ M⁻¹ for Cu(II)-ECG and Cu(II)-EGCG, respectively for the temperature range 21-42 °C with two binding sites. Thermodynamic parameters obtained are suggestive of the involvement of different modes of interaction during binding for each complex although both were found to be intercalating in nature. Circular dichroism studies and variations in the melting temperature reveal unwinding of the ct-DNA helix with conformational changes due to binding.     

Assessment of the anthelmintic activity and toxicity of metabolites of Dalea pogonathera (Fabaceae)

Dalea pogonathera A. Gray was collected as part of a project to determine if metabolites of the genus Dalea have potential for the treatment of human hookworm disease, based on earlier findings of very active materials in D. ornata. We report here the isolation, characterization, and results of ex vivo bioassays of a new chalcone pogonatheridin A (1), and three new prenylated flavanones (3, 6, and 12). The isolated known compounds, a chalcone (2), flavanones (4,5,7-11,13,14), and a flavan-3-ol (15), were also examined. Pogonatheridin A (1) reduced the survival of the adults of Ancylostoma ceylanicum hookworm by 12.5 % (50 μg/mL), while all other compounds showed very weak or no activity. The compounds were tested (50 μg/mL) for toxicity to healthy mammalian cells. Seven of them (2-8) showed > 98 % reduction in survival of splenocytes, while 1 was somewhat less toxic at 74.3 % reduction in survival. While metabolites of D. pogonathera did not show promise as potential anthelmintics for hookworm disease, the toxicity information is of interest, and the rich diversity of metabolites of Dalea spp. remains apparent.