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Phosphoprotein Phosphatase 1 Isoforms Alpha and Gamma Respond Differently to Prodigiosin Treatment and Present Alternative Kinase Targets in Melanoma Cells 在黑色素瘤细胞中,磷酸化磷酸酶1亚型α和γ对Prodigiosin治疗的反应不同,并提出了替代激酶靶点
Pub Date : 2014-04-23 DOI: 10.4236/JBPC.2014.52008
M. Fardilha, J. Figueiredo, M. Espona-Fiedler, Juliana Felgueiras, L. Korrodi-Gregório, S. Esteves, S. Rebelo, O. D. C. E. Silva, E. F. C. E. Silva, R. Pérez-Tomás
Reversible protein phosphorylation is a central regulatory mechanism of cell function. Deregulation of the balanced actions of protein kinases and phosphatases has been frequently associated with several pathological conditions, including cancer. Many studies have already addressed the role of protein kinases misregulation in cancer. However, much less is known about protein phosphatases influence. Phosphoprotein Phosphatase 1 (PPP1) is one of the major serine/threonine protein phosphatases who has three catalytic isoforms: PPP1CA, PPP1CB, and PPP1CC. Its function is achieved by binding to regulatory subunits, known as PPP1-interacting proteins (PIPs), which may prefer a catalytic isoform. Also, some inhibitors/enhancers may exhibit isoform specificity. Here we show that, prodigiosin (PG), a molecule with anticancer properties, promotes the formation of PPP1CA-AKT complex and not of PPP1CC-MAPK complex. Both, AKT and MAPK, are well-known PIPs from two pathways that crosstalk and regulate melanoma cells survival. In addition, the analysis performed using surface plasmon resonance (SPR) technology indicates that PPP1 interacts with obatoclax (OBX), a drug that belongs to the same family of PG. Overall, these results suggest that PG might, at least in part, act through PPP1C/PIPs. Also, this study is pioneer in demonstrating PPP1 isoform-specific modulation by small molecules.
可逆蛋白磷酸化是细胞功能的中心调控机制。蛋白激酶和磷酸酶平衡作用的失调经常与包括癌症在内的几种病理状况有关。许多研究已经解决了蛋白激酶失调在癌症中的作用。然而,对蛋白磷酸酶的影响知之甚少。磷酸蛋白磷酸酶1 (PPP1)是主要的丝氨酸/苏氨酸蛋白磷酸酶之一,有三个催化亚型:PPP1CA、PPP1CB和PPP1CC。它的功能是通过结合调控亚基实现的,被称为ppp1相互作用蛋白(PIPs),这可能更倾向于催化异构体。此外,一些抑制剂/增强剂可能表现出异构体特异性。我们发现,一种具有抗癌特性的分子,能促进PPP1CA-AKT复合物的形成,而不是PPP1CC-MAPK复合物的形成。AKT和MAPK都是众所周知的PIPs,它们通过两条通路相互作用并调节黑色素瘤细胞的存活。此外,使用表面等离子体共振(SPR)技术进行的分析表明,PPP1与obatoclax (OBX)相互作用,OBX是一种属于PG家族的药物。总的来说,这些结果表明PG可能(至少部分)通过PPP1C/PIPs起作用。此外,本研究是小分子对PPP1异构体特异性调节的先驱。
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引用次数: 0
Atherogenic Indices and HDL Particle Size as Laboratory Parameters to Evaluate Cardiovascular Risk in the Presence of Dyslipidemia 在血脂异常的情况下,动脉粥样硬化指数和高密度脂蛋白颗粒大小作为评估心血管风险的实验室参数
Pub Date : 2014-04-23 DOI: 10.4236/JBPC.2014.52004
Ana Paula Caires dos Santos, M. S. Vieira, D. Deus, Rogério Jorge B. de Oliveira, A. Morikawa, Roque Aras Júnior, A. Atta, F. D. Couto, R. Maranhão, R. Couto
Dyslipidemia may influence enzymes and transfer proteins needed to the lipoprotein particle remodeling. Calculated indices and evaluation of lipoprotein particle size have widely been used to predict cardiovascular risk. The aim of this study was to evaluate HDL particle size and LDL particle size estimate based on TG/HDL-C as well as apoB/apoA-I ratio as possible marker and atherogenic indices, respectively, of cardiovascular disease risk in the presence of dyslipidemia. We evaluated 100 individuals of both gender, without treatment with lipid-lowering drugs, 27 normolipidemic and 73 dyslipidemic, such as isolated hypercholesterolemia (n = 16), isolated hypertriglyceridemia (n = 17), low HDL-C (n = 26) and mixed dyslipidemia (n = 14). The HDL particle size did not differ between groups. The TG/HDL-C ratio was higher in groups with isolated hypertriglyceridemia (4.2 ± 1.5), low HDL-C (5.2 ± 3.1) and mixed dyslipidemia (5.3 ± 1.6). The apoB/apoA-I ratio was increased in all groups of dyslipidemia (apoB/apoA-I > 0.5) when compared to normolipidemic (apoB/apoA-I = 0.5, p
血脂异常可能影响脂蛋白颗粒重塑所需的酶和转运蛋白。计算指标和评价脂蛋白颗粒大小已被广泛用于预测心血管风险。本研究的目的是评估基于TG/HDL- c和apoB/apoA-I比值的HDL颗粒大小和LDL颗粒大小,分别作为血脂异常存在时心血管疾病风险的可能标记和动脉粥样硬化指标。我们评估了100名男女,未使用降脂药物治疗,27名正常血脂患者和73名血脂异常患者,如孤立性高胆固醇血症(n = 16),孤立性高甘油三酯血症(n = 17),低HDL-C (n = 26)和混合血脂异常(n = 14)。两组间HDL颗粒大小无差异。孤立性高甘油三酯血症组(4.2±1.5)、低HDL-C组(5.2±3.1)、混合性血脂异常组(5.3±1.6)TG/HDL-C比值较高。与正常血脂组(apoB/apoA-I = 0.5, p)相比,所有血脂异常组(apoB/apoA-I = 0.5)的apoB/apoA-I比值均升高
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引用次数: 4
Gibbs-Donnan Potential as a Tool for Membrane Vesicles Polarization Gibbs-Donnan电位作为膜囊极化的工具
Pub Date : 2014-04-23 DOI: 10.4236/JBPC.2014.52009
Mazur Iuliia, Kosterin Sergiy, V. Tetyana, Shkrabak Oleksandr
It has been theoretically predicted that under conditions leading to Gibbs-Donnan equilibrium in case when size of one compartment is very different from another (as in system “membrane vesicle/liposomes—incubation medium”) stable transmembrane potential can be formed, which value is sufficient to fit requirement of real transmembrane potential. Four partial cases were considered with different location and charge of impermeable ion and it was concluded that locations of impermeable ions in medium provide stable transmembrane potential with sufficient value of 60 - 70 mV. Potential-sensitive probe, such as DiOC6(3) and oxonol VI, were used to confirm the calculated potential. According to the change in fluorescence level and emission/excitation shift, a stable and relatively high transmembrane potential can be formed if salt of impermeable ion is located in incubation medium. Impermeable cations and anions may be used to create positive and negative transmembrane potential respectively.
从理论上预测,在导致Gibbs-Donnan平衡的条件下,当一个腔室的大小与另一个腔室的大小相差很大时(如在“膜泡/脂质体-培养液”系统中),可以形成稳定的跨膜电位,其值足以满足实际跨膜电位的要求。考虑了4种不透离子位置和电荷不同的局部情况,认为介质中不透离子的位置提供了稳定的跨膜电位,其足够值为60 ~ 70 mV。用DiOC6(3)、oxonol VI等电位敏感探针对计算电位进行确认。根据荧光水平的变化和发射/激发位移,在培养介质中加入不渗透离子盐可以形成稳定且较高的跨膜电位。不可渗透的阳离子和阴离子可分别用于产生正的和负的跨膜电位。
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引用次数: 1
Non-Local pH Shifts and Shared Changing Angular Velocity Magnetic Fields: Discrete Energies and the Importance of Point Durations 非局部pH位移和共享变化角速度磁场:离散能量和点持续时间的重要性
Pub Date : 2014-04-23 DOI: 10.4236/JBPC.2014.52006
Nicolas Rouleau, Trevor N. Carniello, M. Persinger
Macroscopic productions of “non-locality” or “excess correlations” of dynamic changes within media between two spaces could be utilized as alternative communication systems. Previous experiments have shown that injections of a weak acid within one of two volumes of spring water sharing the same patterned circular magnetic fields with changing angular accelerations separated by non-traditional (5 m) distances were associated with opposite (basic) shifts in pH within the non-injected, non-local volume. In the present experiments, employing a different technology, pairs of beakers separated by 1 m containing either 25 cc, 50 cc, or 100 cc of spring water were placed within toroids generating weak (30, 300 nT) changing acceleration magnetic fields with 1 ms, 2 ms, or 3 ms point durations or a field whose point durations changed. When a proton source (weak acid) was injected into one beaker (local) pH shifts in the other (non-local) beaker exhibit increased acidity for the 3 ms point duration but increased alkalinity for the 1 ms duration. Neither intermittent point durations nor variable point durations for the same volumes of water placed between the two magnetic field-coupled beakers exhibited significant changes from baseline. Contingent upon the point duration of the applied field, the pH shift was consistent with a fixed quantity of decreased free protons (increased pH) or increased protons (decreased pH) in the non-local beakers. The opposite directions of the pH shifts at 1 ms and 3 ms that correspond to quantitative cosmological solutions for electrons and protons suggest these results may reflect a fundamental physical process.
在两个空间之间的媒介中动态变化的“非局域性”或“过度相关性”的宏观生产可以用作替代通信系统。先前的实验表明,在两卷泉水中的一卷中注入弱酸,这些泉水具有相同的圆形磁场,并且以非传统(5米)距离分隔角加速度变化,在未注入的非局部体积中,pH值会发生相反的(基本的)变化。在目前的实验中,采用了一种不同的技术,将含有25cc, 50cc或100cc泉水的一对烧杯放置在环形管内,产生微弱(30,300 nT)变化的加速磁场,其点持续时间分别为1ms, 2ms或3ms,或点持续时间发生变化的场。当质子源(弱酸)注入一个烧杯(局部)时,另一个烧杯(非局部)的pH值变化在3ms点持续时间内表现出酸性增加,但在1ms持续时间内碱性增加。对于放置在两个磁场耦合烧杯之间的相同体积的水,间歇点持续时间和可变点持续时间都没有显示出与基线相比的显着变化。根据施加电场的点持续时间,pH值的变化与非局部烧杯中固定数量的自由质子减少(pH值增加)或质子增加(pH值降低)一致。对应于电子和质子的定量宇宙学解的1 ms和3 ms的pH位移方向相反,表明这些结果可能反映了一个基本的物理过程。
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引用次数: 10
Genetic Variation in Salamandra infraimmaculata from Different Habitats Using Amplified Fragment Length Polymorphism 利用扩增片段长度多态性分析不同生境下小鲵的遗传变异
Pub Date : 2014-04-23 DOI: 10.4236/JBPC.2014.52007
G. Degani, T. Goldberg, E. Nevo
The purpose of the present study was to examine the genetic variation in Salamandra infraimmaculata from different breeding site habitats using the Amplified Fragment Length Polymorphism (AFLP) method. The results of the dendogram from a hierarchical cluster analysis show that the grouping of S. infraimmaculata as cluster 5 differs from all the other clusters, including the St1 (Tel-Dan stream) population, which was the most predictable. Five Haplogroups (Hg) were characterized. The mean number of alleles per locus in each population (Ne) ranged from 10.566 (Sp1) to 2.720 (Po6). An average estimated heterozygosity (He) by population ranged from 0.100 (Po6) to 0.186 (St1). Population St1, a permanent breeding site where water was available all year round, exhibited the highest level of polymorphism, while population Po6, from the ephemeral breeding site, exhibited the lowest level of polymorphism. Gene flow between clusters showed that clusters 3 and 4 are sources of migrants and also receive gene flow, while clusters 1 and 2 may be a source of migrants but may not receive much gene flow. A phylogenetic analysis, based on clustering using Nei’s genetic distance, demonstrated that the Tel-Dan population is located on a separate branch within its sub-population. The conclusion of the present study shows that the genetic divergence among isolated populations is not correlated to distance but is affected by the variation of habitats.
本研究采用扩增片段长度多态性(AFLP)方法,对不同孳生地的小红花鲵的遗传变异进行了分析。分层聚类分析的树图结果表明,S. infraimmaculata的聚类与其他所有聚类不同,包括最具可预测性的St1 (Tel-Dan stream)种群。测定了5个单倍群(Hg)。每个群体每个位点的平均等位基因数(Ne)在10.566 (Sp1) ~ 2.720 (Po6)之间。群体平均杂合度(He)估计范围为0.100 (Po6) ~ 0.186 (St1)。常年有水的永久繁殖地种群St1的多态性水平最高,而短暂繁殖地种群Po6的多态性水平最低。集群间的基因流动表明,集群3和集群4是移民的来源,也接受基因流,而集群1和集群2可能是移民的来源,但可能接受的基因流不多。基于Nei遗传距离聚类的系统发育分析表明,Tel-Dan种群位于其亚种群的一个单独分支上。本研究的结论表明,孤立居群间的遗传分化与距离无关,而受生境变化的影响。
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引用次数: 5
Hemin Protects U937 Cells from Oxidative Stress via Glutathione Synthesis 血红蛋白通过谷胱甘肽合成保护U937细胞免受氧化应激
Pub Date : 2014-04-23 DOI: 10.4236/JBPC.2014.52003
Y. Satoh, Kyohei Oyama, K. Sakurai
Although heme oxygenase-1 and glutathione play important roles in the antioxidant defense system, the sharing and/or cross-talking of the HO-1 and GSH system remain poorly understood. The object of this study is to determine whether the glutathione system is involved in the antioxidant function of hemin. Hydrogen peroxide decreased the viability of the human leukemic monocyte lymphoma cell line U937. When these cells were pretreated with hemin before the addition of hydrogen peroxide, cell death was prevented. An inhibitor of heme oxygenase-1 or glutathione biosynthesis significantly abolished this protective effect of hemin. These results suggest that both heme oxygenase-1 and glutathione are involved in the protective effects of hemin against U937 cell death, which was induced by hydrogen peroxide. Hemin induced an increase in glutathione levels following the upregulation of the gene expression and protein levels of glutamate-cysteine ligase catalytic subunit. The inhibitor of heme oxigenase-1 inhibited the upregulation of glutamate-cysteine ligase catalytic subunit expression. These results suggest that hemin induces glutathione synthesis through heme oxygenase-1 activation to protect cells from hydrogen peroxide-induced oxidative stress.
虽然血红素加氧酶-1和谷胱甘肽在抗氧化防御系统中发挥重要作用,但HO-1和谷胱甘肽系统的共享和/或相互作用仍然知之甚少。本研究的目的是确定谷胱甘肽系统是否参与血红蛋白的抗氧化功能。过氧化氢降低人白血病单核细胞淋巴瘤细胞系U937的活力。当这些细胞在加入过氧化氢之前用血红蛋白预处理时,细胞死亡被阻止。血红素加氧酶-1或谷胱甘肽生物合成抑制剂显著地消除了血红素的这种保护作用。提示血红素加氧酶-1和谷胱甘肽均参与血红素对过氧化氢诱导的U937细胞死亡的保护作用。在谷氨酸-半胱氨酸连接酶催化亚基基因表达和蛋白水平上调后,Hemin诱导谷胱甘肽水平升高。血红素氧化酶-1抑制剂抑制谷氨酸-半胱氨酸连接酶催化亚基表达上调。这些结果表明,血红素通过激活血红素氧化酶-1诱导谷胱甘肽合成,保护细胞免受过氧化氢诱导的氧化应激。
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引用次数: 0
A Unique Conformational Behaviour of Glutamine Peptides 谷氨酰胺肽的独特构象行为
Pub Date : 2014-04-23 DOI: 10.4236/JBPC.2014.52005
M. Baskar, M. Shafique, F. S. Nandel
Trinucleotide repeat expansions (CAG) lead to increase in glutamine residues and hence increase in glutamine stretch. This leads to number of neurodegenerative diseases. Therefore, the conformation of poly Q of varying chain lengths has been investigated by quantum mechanical and molecular dynamics approaches. Glutamine contains amide linkage in the side chain. It is the interaction between side chain amide linkage and the peptide bond of the backbone which dictates the conformational behaviour. Some of the glutamine residues adopt phi psi values corresponding to poly-L-proline type II structure. Not more than three glutamine residues are found to have the same set of values and hence polyglutamine is adopting random coil structure. Carbonyl-carbonyl, CH-O interactions and hydrogen bond formation involving backbone and side chain amide chain linkages are found to contribute to the stability of the adopted structure. In simulation studies due to interaction of water molecules with the amide linkages the values undergo change and this leads to weakening of carbonyl-carbonyl interactions and hydrogen bonds. The conformational behaviour of polyglutamine peptides is shown to be chain length dependent and this may provide some insight regarding the aggregation behaviour of proteins containing poly Q stretch. Possibly this is the first systematic study of the conformational behaviour of polyglutamine peptides.
三核苷酸重复扩增(CAG)导致谷氨酰胺残基增加,从而增加谷氨酰胺拉伸。这导致了许多神经退行性疾病。因此,采用量子力学和分子动力学的方法研究了不同链长的聚Q的构象。谷氨酰胺在侧链中含有酰胺键。侧链酰胺键与主链肽键之间的相互作用决定了其构象行为。一些谷氨酰胺残基采用与聚l -脯氨酸II型结构相对应的phi - psi值。发现不超过三个谷氨酰胺残基具有相同的一组值,因此多谷氨酰胺采用随机线圈结构。羰基-羰基、CH-O相互作用以及涉及主链和侧链酰胺链连接的氢键形成有助于所采用结构的稳定性。在模拟研究中,由于水分子与酰胺键的相互作用,这些值发生了变化,这导致羰基-羰基相互作用和氢键的减弱。聚谷氨酰胺肽的构象行为被证明是链长依赖的,这可能为含有聚Q拉伸的蛋白质的聚集行为提供一些见解。这可能是第一次系统地研究聚谷氨酰胺肽的构象行为。
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引用次数: 0
Geometrical criteria for left-handed twists within protein beta-strands 蛋白质-链内左旋扭曲的几何准则
Pub Date : 2014-01-15 DOI: 10.4236/JBPC.2014.51002
B. Caudron, J. Jestin
Using a statistical analysis on beta-sheet structures from the Protein Data Bank, characteristic angles within beta-strands were correlated to the nature of the side chains. The twists were computed from the atomic coordinates of five consecutive amino acids’ alpha carbons from single beta-strand sequences. Conditions on the angles for twists to be mainly left-handed are given together with the frequency of occurrence for these non-standard geometrical properties within protein beta-strands. Applications in protein structure prediction and CASP challenges in particular are envisioned by making use of the probabilities of occurrence in protein structures of angle value ranges for given amino acids.
通过对来自蛋白质数据库的β -片结构的统计分析,β -链内的特征角度与侧链的性质相关。这些扭曲是根据单个-链序列中五个连续氨基酸α碳的原子坐标计算出来的。给出了扭曲角度主要为左旋的条件,以及这些非标准几何性质在蛋白质-链中出现的频率。通过利用给定氨基酸的角度值范围内蛋白质结构的发生概率,设想了在蛋白质结构预测和CASP挑战中的应用。
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引用次数: 1
Antioxidants as effective remedies at hepatotoxic action of carbon tetrachloride 抗氧化剂对四氯化碳肝毒性作用的有效补救
Pub Date : 2014-01-15 DOI: 10.4236/JBPC.2014.51001
K. Hovnanyan, V. Mamikonyan, A. Margaryan, K. Sargsyan, M. Hovnanyan, M. Karagyozyan, K. Karageuzyan
The liver, as the central metabolic organ, plays an important role in metabolism of carbohydrates, lipids and proteins. In the environment, there are many hepatotoxic xenobiotics. CCl4 is a free radical toxic for organelles of hepatocytes. The presented work was studying the character of ultrastructural changes in CCl4-intoxicated hepatocytes as well as the tissue after treatment by grape seed extract (GSE), α-tocopherol (α-T). As a result of cell bioremediation effect and reactions against corruptive factors, the liver has a high regeneration ability.
肝脏作为中枢代谢器官,在碳水化合物、脂质和蛋白质的代谢中起着重要作用。在环境中,有许多肝毒性的外源性药物。CCl4是一种对肝细胞细胞器有毒性的自由基。研究葡萄籽提取物(GSE)、α-生育酚(α-T)对ccl4中毒大鼠肝细胞及肝组织超微结构的影响。由于细胞的生物修复作用和对腐败因素的反应,肝脏具有很高的再生能力。
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引用次数: 7
Morphology observation of dipalmitoyl phosphatidyl choline (DPPC) monolayer on water surface by dropping method 水滴法观察水表面双棕榈酰磷脂酰胆碱(DPPC)单分子膜的形态
Pub Date : 2013-11-21 DOI: 10.4236/JBPC.2013.44016
D. Yoshida, T. Yokoyama, T. Shimoaki, T. Tomita, T. Yoshida, Yasushi Yamamoto, K. Taga, Ayumi Sumino, Takehisa Dewa, M. Nango, Masato Yamamoto, Zameer Sheravani
Preparation of DPPC lipid monolayer in water trough has been done by dropping method and compared with compression method. Monolayer was studied by surface pressure isotherm, fluorescence microscopy, Brewster angle microscopy, and infrared external reflection spectroscopy. Results of these measurements showed that dropping method gave better results compared to compression method. In dropping method, transition from liquid expanded state to liquid condensed is gradual compared to sharp one in compressed method. During monolayer formation, adjustment and interaction between hydrophilic part of lipid and water and among hydrophobic part of lipid molecule are slow, stable, and more natural as worked out from surface area versus pressure isotherm. At a given molecular area, surface pressure is less compared to compression method thus monolayer is in more fluidic state in dropping method than compression method. The observation was supported by all techniques described above.
采用滴落法在水槽中制备DPPC脂质单层,并与压缩法进行了比较。通过表面压力等温线、荧光显微镜、布鲁斯特角显微镜和红外外反射光谱对单层膜进行了研究。结果表明,与压缩法相比,滴法的效果更好。在滴法中,从液体膨胀状态到液体冷凝状态的转变是渐进的,而在压缩法中则是急剧的。在单层形成过程中,脂质亲水部分与水之间、疏水部分与脂质分子之间的调节和相互作用是缓慢、稳定和自然的,这是由表面积与压力等温线得出的。在给定的分子面积上,表面压力比压缩法小,因此在滴法中单层比压缩法更处于流态。上述所有技术都支持这一观察结果。
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引用次数: 11
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生物物理化学(英文)
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