Pushing, pulling and trapping--modes of motor protein supported protein translocation

Protein translocation across the cellular membranes is an ubiquitous and crucial activity of cells. This process is mediated by translocases that consist of a protein conducting channel and an associated motor protein. Motor proteins interact with protein substrates and utilize the free energy of ATP binding and hydrolysis for protein unfolding, translocation and unbinding. Since motor proteins are found either at the cis- or trans-side of the membrane, different mechanisms for translocation have been proposed. In the Power stroke model, cis-acting motors are thought to push, while trans-motors pull on the substrate protein during translocation. In the Brownian ratchet model, translocation occurs by diffusion of the unfolded polypeptide through the translocation pore while directionality is achieved by trapping and refolding. Recent insights in the structure and function of the molecular motors suggest that different mechanisms can be employed simultaneously.     

汉防己甲素联合顺铂对乳腺癌细胞的作用

目的:汉防己甲素和顺铂联合作用乳腺癌细胞,来提高癌细胞对顺铂的敏感性.方法:运用原子力显微镜对乳腺癌细胞表面的超微结构进行表征;MTr法和流式细胞术检测细胞的生长抑制率和细胞周期变化.结果:顺铂和汉防己甲素分别作用乳腺癌细胞48h IC50值为26.33μmoL/l和5.6μmok/l;联合用药组的IC50值为汉防己甲素2.5μmol/L和顺铂13.32μmol/l,在低浓度的联合用药作用乳腺癌细胞24h细胞膜表面结构被破坏,产生孔洞,作用48h被严重破坏使细胞周期在S比例增加为51.7%±0.30%.结论:提高了癌细胞对抗癌药物的敏感性,联合用药通过改变了细胞膜的结构对癌细胞进行有效杀伤,抑制肿瘤细胞生长.     

Poly(dG)-poly(dC) DNA appears shorter than poly(dA)-poly(dT) and possibly adopts an A-related conformation on a mica surface under ambient conditions

Three types of DNA: approximately 2700 bp polydeoxyguanylic olydeoxycytidylic acid [poly(dG)-poly(dC)], approximately 2700 bp polydeoxyadenylic polydeoxythymidylic acid [poly(dA)-poly(dT)] and 2686 bp linear plasmid pUC19 were deposited on a mica surface and imaged by atomic force microscopy. Contour length measurements show that the average length of poly(dG)-poly(dC) is approximately 30% shorter than that of poly(dA)-poly(dT) and the plasmid. This led us to suggest that individual poly(dG)-poly(dC) molecules are immobilized on mica under ambient conditions in a form which is likely related to the A-form of DNA in contrast to poly(dA)-poly(dT) and random sequence DNA which are immobilized in a form that is related to the DNA B-form.     

Sexual dimorphism in rat left atrial function and response to adrenergic stimulation

A number of investigations in humans and animals suggest that there may be intrinsic sex-associated differences in cardiac function. Using left atrial preparations from male and female rat hearts, we examined differences in myocardial function and response to adrenergic agonists. Contractile parameters were measured in isolated atria by conventional isometric methods in the absence or presence of isoproterenol or phenylephrine. Responsiveness to Ca²⁺ was measured in detergent-skinned atrial fibers and actomyosin ATPase activity was measured in isolated myofibrils. Tetanic contractions were generated by treating the atrium with ryanodine followed by high frequency stimulation. Developed force was greater and maximal rates of contraction and relaxation were more rapid in the female atrium. The relationship between Ca²⁺ concentration and force in both intact atria and detergent-skinned atrial fibers in females fell to the left of that for males. At low Ca²⁺ concentrations, skinned fibers from female atria generated more force and myofibrils from female atria had higher myosin ATPase activity than males. Tetanic contraction in the presence of high extracellular Ca²⁺ was greater in female atria. Male atrium had larger inotropic responses to isoproterenol and to phenylephrine, but drug-elicited cAMP and inositol phosphate production did not differ between sexes. The results demonstrate sex-related differences in atrial function that can be partially explained by greater myofibrillar Ca²⁺-sensitivity in females. A potential contribution of sarcolemmal Ca2+ influx is suggested by greater tetanic contraction in ryanodine-treated female atrium. The larger response of males to adrenergic stimulation does not appear to be explained by higher production of relevant second messengers. Future studies will investigate the role of sex hormones in these sexually dimorphic responses and may indicate a need for gender-specific therapeutic interventions for myocardial dysfunction.     

A hidden aggregation‐prone structure in the heart of hypoxia inducible factor prolyl hydroxylase

Prolyl hydroxylase domain‐containing protein 2 (PHD2), as one of the most important regulators of angiogenesis and metastasis of cancer cells, is a promising target for cancer therapy drug design. Progressive studies imply that abnormality in PHD2 function may be due to misfolding. Therefore, study of the PHD2 unfolding pathway paves the way for a better understanding of the influence of PHD2 mutations and cancer cell metabolites on the protein folding pathway. We study the unfolding of the PHD2 catalytic domain using differential scanning calorimetry (DSC), fluorescence spectroscopy, and discrete molecular dynamics simulations (DMD). Using computational and experimental techniques, we find that PHD2 undergoes four transitions along the thermal unfolding pathway. To illustrate PHD2 unfolding events in atomic detail, we utilize DMD simulations. Analysis of computational results indicates an intermediate species in the PHD2 unfolding pathway that may enhance aggregation propensity, explaining mutation‐independent PHD2 malfunction. Proteins 2016; 84:611–623. © 2016 Wiley Periodicals, Inc.     

一种基于单分子纳米操纵的有序化测序策略

尽管包括人类在内的许多生物物种的基因组测序工作已经完成,但由于现有测序技术的限制,大部分复杂基因组还存在很多大大小小的缺口.缺口的填补以及对其他重复序列区域的测序迫切需要全新的思路和技术.基于在DNA单分子定位切割和拾取方面的实验进展,提出了一种基于原子力显微镜纳米操纵技术的单分子有序化测序策略.计算机模拟的结果表明,这一方法和策略是可行的,有助于解决目前测序工作中所遇到的一些棘手问题.     

人外周血单个核细胞与脐带间充质干细胞共培养的AFM研究

采用原子力显微镜与倒置显微镜在细胞层次上观察了人外周单个核细胞(PBMCs)与同种异源脐带间充质干细胞(hUC-MSCs)共培养的过程,并在单细胞水平上分析了共培养前后人外周单个核细胞的形貌和生物物理性质。结果发现:共培养后贴壁人外周单个核细胞的形态发生了很大的改变,并且表面分布着大小不一的颗粒状聚合物。利用AFM高空间分辨的力位移曲线测量系统,发现共培养72h后培养上清中人外周单个核细胞、贴壁的人外周单个核细胞的粘滞力分别是单纯培养72h的人外周单个核细胞的2倍、5倍,而细胞的硬度分别是单纯培养人外周单个核细胞的1.5倍、2倍。CCK-8检测提示,共培养过程中,干细胞的生长与外周血单个核细胞的生长出现了竞争作用。通过AFM探测人外周单个核细胞与脐带间充质干细胞共培养的可视化数据,有助于更好地了解间充质干细胞与外周血单个核细胞的相互作用。     

基于生物适应进化原理论证生物进化的动力

文章从现有主流生物进化理论存在的问题入手, 以生物适应进化原理为认识基础, 讨论生物进化的动力, 以求对生物进化机制有一个新的认识。在薛定谔“生命赖负熵生存”观点的指导下, 提出了“负熵流”包括能量流、物质流和信息流, 以及负熵流是生命生存和发育的动力的观点。作者在原有生物适应进化原理基础上, 修改完善并提出了“DNA、RNA和蛋白质在环境作用下的生物适应进化调控系统”理论, 并根据系统发育是个体发育的“积分”的观点, 推论得出生物与环境的负熵差引起的负熵流也是生命进化的动力, 对生物进化机制作出了新的理解。基于这样的生物进化机制的认识, 提出了“进化是一个子系统在其上一等级系统中, 将自身全部或部分信息遗传给下一代子系统, 并在其适应上一等级系统过程中, 产生一些新质, 终止一些旧质, 从而在其上一等级系统中得以延续的变化过程”的概念, 并探讨了一些与进化有关的其他争议问题。     

城市化区域生态风险驱动力及管控策略——以北京市为例

快速城市化导致城市及周边区域生态风险不断增加,认知城市化区域生态风险,探究其变化的驱动因素,提出行之有效的风险管控策略,对城市化区域生态环境与社会经济协调发展具有重要意义。以北京市为例,基于生态系统服务对生态风险进行了定量表征,分析了城市化因素对生态风险的驱动作用,探究了生态风险管控策略。研究结果表明：不透水地表扩张、经济增长、人口增长均对生态风险有显著的驱动作用,其中不透水地表扩张的驱动作用占主导地位。经济增长和人口增长通过影响不透水地表率间接作用于生态风险,在2005年时,人口密度对生态风险的驱动作用大于GDP强度,在2010年及之后,GDP强度对生态风险的驱动作用则变得比人口密度的驱动作用更大。北京市的生态风险管控策略为：在不透水地表率介于40%—70%的区域降低不透水地表率和实施土地利用及景观格局优化;不透水地表率大于70%的区域应防止不透水地表进一步扩张;不透水地表率低于40%的区域需通过提升生态系统质量、强化风险防范、降低风险暴露等途径来管控风险。此外,应通过提高海淀、昌平、通州、顺义等辖区的人口和经济城市化集约水平来降低生态风险。本研究阐明了城市化对区域生态风险的驱动作用...