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大紫丹参的多酚类化合物 总被引:9,自引:0,他引:9
从云南丽江产大紫丹参( Salvia przewalskii Maxim .) 的根部分离得到11 个多酚类化合物, 其中8 个鉴定为已知的原儿茶醛, 原儿茶酸, 咖啡酸, R- ( + ) - β- D- (3 , 4 - 二羟基苯基) - 乳酸, 迷迭香酸, 迷迭香酸甲酯, 紫草酸和紫草酸B。另外3 个为紫草酸B的甲酯化衍生物, 即紫草酸B二甲酯, 9″- 紫草酸B 单甲酯和9- 紫草酸B 甲单酯。它们的结构通过波谱方法得到鉴定。研究结果表明, 大紫丹参含有与正品丹参相似的酚类化合物。 相似文献
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蜂毒突变体对紫膜质子泵功能的影响 总被引:1,自引:0,他引:1
具有光驱动质子泵功能的嗜盐菌紫膜是一种被广泛研究的生物膜系统。利用毫秒级闪光动力学谱仪研究蜂毒不同突变体对紫膜质子泵功能的影响。实验采用对硝基苯酚(p-nitrophenol)作为PH敏感染料来研究紫膜蛋白-菌紫质(Bacteriorhodopsin,简称)的光反应和质子泵功能。在初步的实验中发现,在一定的温度范围内,光循环过程随着温度的升高而加快,质子泵功能却基本保持不变。另外,由于蜂毒突变体具有不同的插膜特性和不同的电荷量,通过比较它们对BR质子泵功能的影响,揭示了蜂毒小肽C端和N端的不同带电状态对质子泵功能所起的作用不同。实验结果有力地证实了蜂毒或其突变体与菌紫质蛋白具有直接的相互作用,这种相互作用的强弱与蜂毒突变体电荷的多少密切相关 相似文献
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本文研究了分别从红藻多管藻(Polysiphoniaurceolate)和条斑紫菜(Porphyrayezoensis)中提取的两种不同光谱类型的R-藻红蛋白R-phycoerythrin激发强度相关的皮秒(10-12秒)荧光衰减动力学过程。结果发现:随激发光强增大,单重态-单重态激子湮灭发生(其衰减过程约为60~80皮秒),并引起荧光量子产率下降。这两种B-藻红蛋白在相同光强激发下,表现出不同的单重态一单重态激子湮灭过程,主要因它们处于激发态的发色团数目不同所致。 相似文献
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RRM RNA 结合蛋白在基因的转录后调控中起着重要作用.人GRY-RBP 是我们实验室最近克隆的一个新的全长cDNA,编码一个RRM RNA 结合蛋白.GRY-RBP 的全编码区用PCR扩增后,克隆到pET30a+ 表达载体中,在E.coli中获得了高效表达,表达的GRY-RBP重组蛋白占细菌总蛋白的21% .利用融合在GRY-RBPN 端的His.Tag,采取亲和层析的方法纯化了表达的重组蛋白.为了检测GRY-RBP的RNA 结合活性及其所结合的RNA 性质,将表达的蛋白与poly(A)-Sepharose 4B结合,发现GRY-RBP能与polyA (A)结合,结合活性能被可溶性的poly(A)、poly(C)减弱.改变NaCl浓度和加入不同浓度的酵母tRNA竞争物后,poly(A)结合活性不变. 相似文献
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从青岛采集的多管藻(Polysiphoniaurceolata)中分离得到的R-藻蓝蛋白,在pH=7.0的0.05mol/L磷酸盐-硫酸铵缓冲液中,使用悬滴气相扩散法获得适合X光衍射分析用单晶。经Buerger徘循照相和XRD—100面探测仪分析,R-藻蓝蛋白晶体属于四方晶系,空间群为P41(3)212,晶胞参数:a=b=137.5c=218.5α=β=γ=90°。用等比重梯度柱法测定了晶体和母液的比重分别为1.19和1.09。根据分子量与晶胞体积估算,一个不对称单位含有一个分子,推测它的分子聚集态形式为(αβ)3。 相似文献
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本文研究用非离子表面活性剂Triton X-100处理后的细菌视紫红质(BR Bacteriorhodo-psin)光循环中间产物M_(412)动力学过程的变化.实验结果表明,用不同浓度的Triton处理pH=6.5的BR体系时,其中间产物M_(412).快衰减成分的半衰期(τ_(1/2)~f)在Triton浓度为0.05%(w/w)附近突然变慢,随着Triton浓度的加大,τ_(1/2)~f又逐渐加快;慢衰减部分的半衰期(τ_(1/2)~s)则随Tri-ton浓度的增加逐渐变慢.BR的生色团峰发生蓝移.说明不同浓度的Triton在水溶液中聚集状态不同,可不同程度地破坏膜脂的液晶态结构,从而导致镶嵌在其中的BR发生构象的变化,使转运质子的氢键通道受到不同程度的影响,故质子泵转运通道发生改变、致使M_(412)的衰减速率改变. 相似文献
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We report that Triton X-100 can alter the temporal sequence of the light-induced proton uptake and release of archaerhodopsin 4 (AR4), a proton pumping protein in a species of Halobacteria from a Tibetan salt lake. Under physiological conditions, AR4 isolated from the bacterium exhibits a reversed temporal order of proton release and uptake compared to what is observed for bacteriorhodopsin (BR). However, in the presence of Triton X-100 early proton release was observed in AR4 at neutral pH by us. Further, this temporal order for light-driven proton release and uptake for AR4 was found to be recovered after the removal of Triton X-100 by Biobeads. This phenomenon of detergent-induced alteration of the order of proton release and uptake has not yet been reported in any other retinal-containing membrane protein such as BR. Our findings indicate that the function of AR4 is influenced by its self-assembled state, and meanwhile imply some subtle protein-lipid interactions or protein-protein interactions in adjusting the proton pumping behavior of AR4. 相似文献
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Purification of bacteriorhodopsin and characterization of mature and partially processed forms 总被引:2,自引:0,他引:2
L J Miercke P E Ross R M Stroud E A Dratz 《The Journal of biological chemistry》1989,264(13):7531-7535
Bacteriorhodopsin (BR) essentially free of native lipids has been prepared in a highly stable state. Purple membrane was solubilized in Triton X-100 and BR was purified by size exclusion chromatography using 3-[cholamidopropyl)dimethylammonio]-2-hydroxyl-1-propanesulfonic acid (CHAPSO) detergent at pH 5. Molar ratios of phospholipid/BR ranged from 0.4 to 0.05 corresponding to 94-98% phospholipid removal. Purified BR has an absorbance ratio (A280nm/A548nm) of 1.5-1.6 in the dark-adapted state which is the highest purified BR/protein ratio reported to date. The purified BR in CHAPSO shows maximum stability in the pH range 5.0-5.5. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles of native purple membrane and solubilized BR from most Halobacterium halobium JW-3 cultures show 3 higher molecular weight bands in addition to BR. Immunological staining and amino acid sequencing indicates that these additional proteins are partially processed forms of the BR precursor protein. The BR preprotein contains 13 additional amino acids on the NH2 terminus which are removed by post-translational processing in at least four steps. Isoelectric focusing separated most delipidated and non-delipidated BR samples into 8 bands. Incomplete BR post-translational processing BR is thought to be largely responsible for the multiplicity of isoelectric BR species. The principal components have pI values of 5.20 and 5.24 and both have absorption maxima at 550 nm, characteristic of detergent-solubilized BR. BR in Triton X-100 or nonylglucoside, delipidated BR in CHAPSO, and BR in intact purple membrane all have a dark-adapted ratio of 13-cis to all-trans-retinal of 1.9:1. 相似文献
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Assembly of Trp1 in a signaling complex associated with caveolin-scaffolding lipid raft domains 总被引:12,自引:0,他引:12
Lockwich TP Liu X Singh BB Jadlowiec J Weiland S Ambudkar IS 《The Journal of biological chemistry》2000,275(16):11934-11942
Trp1 has been proposed as a component of the store-operated Ca(2+) entry (SOC) channel. However, neither the molecular mechanism of SOC nor the role of Trp in this process is yet understood. We have examined possible molecular interactions involved in the regulation of SOC and Trp1 and report here for the first time that Trp1 is assembled in signaling complex associated with caveolin-scaffolding lipid raft domains. Endogenous hTrp1 and caveolin-1 were present in low density fractions of Triton X-100-extracted human submandibular gland cell membranes. Depletion of plasma membrane cholesterol increased Triton X-100 solubility of Trp1 and inhibited carbachol-stimulated Ca(2+) signaling. Importantly, thapsigargin stimulated Ca(2+) influx, but not internal Ca(2+) release, and inositol 1,4,5-triphosphate (IP(3))-stimulated I(soc) were also attenuated. Furthermore, both anti-Trp1 and anti-caveolin-1 antibodies co-immunoprecipitated hTrp1, caveolin-1, Galpha(q/11), and IP(3) receptor-type 3 (IP(3)R3). These results demonstrate that caveolar microdomains provide a scaffold for (i) assembly of key Ca(2+) signaling proteins into a complex and (ii) coordination of the molecular interactions leading to the activation of SOC. Importantly, we have shown that Trp1 is also localized in this microdomain where it interacts with one or more components of this complex, including IP(3)R3. This finding is potentially important in elucidating the physiological function of Trp. 相似文献
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This study reports the isolation and characterization of a Triton X-100-resistant membrane fraction from homogenates of rod outer segment (ROS) disk membranes purified free of the surrounding plasma membrane. A portion of the ROS disk membrane was found to be resistant to Triton X-100 extraction at 4 degrees C. This detergent-resistant fraction was isolated as a low buoyant density band on sucrose density gradients and exhibited an increase in light scattering detected at 600 nm. Biochemical analysis of the Triton X-100-resistant fraction showed it to be enriched in cholesterol and sphingomyelin relative to phospholipid and in phospholipid relative to protein compared with the soluble fraction. The Triton X-100-resistant membranes described herein did not arise simply from partial solubilization of the ROS disk membranes because detergent-treated low buoyant density fractions isolated from homogenates with octyl glucopyranoside had cholesterol and sphingomyelin content indistinguishable from that of solubilized ROS disk homogenates. Analysis of proteins associated with the Triton X-100-resistant fraction showed it to be enriched in the rim-specific protein ROM-1 and caveolin; surprisingly, the fusion protein peripherin/rds (where rds is retinal degeneration slow), also localized to the disk rim, was entirely absent from the membrane raft domain. The lipid profiles of the Triton X-100-resistant membranes were virtually identical in preparations homogenized in either the light or dark. Slightly more ROM-1 was recovered from samples prepared in the light (23%) than from samples prepared in the dark (13%), but peripherin/rds could not be detected in either preparation. When the Triton X-100-resistant membranes were treated with methyl-beta-cyclodextran to deplete membrane cholesterol, the resultant membranes contained slightly lower levels of ROM-1, specifically in the dimeric form. Cholesterol depletion also resulted in the collapse of the large caveolin complex to monomeric caveolae. The results presented herein characterize a pool of ROM-1, a photoreceptor tetraspanin protein, that may play a regulatory role in peripherin/rds-dependent fusion. 相似文献
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(31)~P—NMR实验表明,小的超声囊受表面活性剂Tween80或TritonX—100作用后磷脂头部基团的运动受到限制,可能与磷脂的磷酸根和水化的表面活性剂分子通过氢键的相互作用有关。多层脂质体加Triton X—100后显示各向同性运动,提示表面活性剂可诱发双层结构的改变。 相似文献
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The effect of Triton X-100 on catalytic properties of acetylcholinesterase from human erythrocytes under acetylcholine hydrolysis, on sensitivity of acetylcholinesterase to specific phosphoorganic inhibitors and eserine, and on the mobility and isoenyme spectrum under analytical electrophoresis in polyacrylamide gel is investigated. Triton X-100, independently on its concentration within 0.05-1.0%, slightly changes V and [S]opt values and increases Km value in 2-3 times. The inhibitory effect of Triton X-100 is mainly competitive, 0.5% Triton X-100 decreases bimolecular constant (kII) of the interaction of acetylcholinesterase with phosphoorganic inhibitor and eserine in 2.5-4 times. In the presence of phosphoorganic inhibitor, kII sharply decreased when 0.02% Triton X-100 was added, and then it did not change under the increase of Triton X-100 concentration up to 1.0%. On the basis of these data, an analytical method of estimating Triton X-100 content in protein solution is proposed. The introduction of 0.1% Triton X-100 into polyacrylamide gel results in considerable quantitative redistribution of acetylcholinesterase isoenzyme fractions and in the change of the mobility of one fraction under electrophoresis. 相似文献
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杨万年 《分子细胞生物学报》1986,(3)
本文研究了蚕豆叶肉原生质体经透明质酸酶、核糖核酸酶、神经氨酸酶、碱性磷酸酶、胰蛋白酶、脂肪酶六种水解酶和SDS、Triton X-100、CTMAB三种表面活性剂以及秋水仙素、细胞松驰素B处理后的电融合过程。结果表明:胰蛋白酶处理后的原生质体融合率明显下降;碱性磷酸酶、脂肪酶以及核糖核酸酶、透明质酸酶、神经氨酸酶处理的原生质体电融合率均有不同程度的上升。Triton X-100和CTMAB促进原生质体的电融合,但较高浓度(0.01%)的SDS起抑制作用。秋水仙素和细胞松驰素B处理的原生质体其电融合率有较大幅度的增高。 相似文献