Incorporation of the transmembrane hydrophobic domain of glycophorin into small unilamellar phospholipid vesicles Ion Flux studies

Human erythrocyte glycophorin is one of the best characterized integral membrane proteins. Reconstitution of the membrane-spanning hydrophobic segment of glycophorin (the tryptic insoluble peptide released when glycophorin is treated with trypsin) with liposomes results in the production of freeze-fracture intrabilayer particles of 80 Å diameter (Segrest, J.P., Gulik-Krzywicki, T. and Sardet, C. (1974) Proc. Natl. Acad. Sci. U.S.A. 71, 3294–3298), with particles appearing at or above a tryptic insoluble peptide concentration of 4 mmol per mol phosphatidylcholine. In the present study, increasing concentrations of tryptic insoluble peptide were added to sonicated small unilamellar egg phosphatidylcholine vesicles and the rate of efflux of ²²Na⁺ was examined by rapid (30 s) gel filtration on Sephadex G-50. Below a concentation of 3–5 mmol tryptic insoluble peptide/mol phosphatidylcholine, ²²Na⁺ efflux occurs at a constant slow rate at given tryptic insoluble peptide concentrations. Above a concentration of 3–5 mM, the rate of efflux is biphasic at given tryptic insoluble peptide concentrations, exhibiting both an initial fast and a subsequent slow component. On the basis of graphic and computer curve-fitting analysis, with increasing tryptic insoluble peptide concentration, the rate of the slow component reaches a plateau at a tryptic insoluble peptide concentration of 3–5 mM and remains essentially constant until much higher concentrations are reached; the fast component increases linearly with increasing tryptic insoluble peptide concentration well beyond 5 mM. The most consistent interpretation of this data is as follows. The slow ²²Na⁺ efflux component is due to perturbations of small unilamellar vesicle integrity by tryptic insoluble peptide monomers. At a tryptic insoluble peptide concentration of 3–5 mmol/mol, a critical concentration is reached following which there is intrabilayer tryptic insoluble peptide self-association. The fast ²²Na⁺ efflux component is due to the increasing presence of tryptic insoluble peptide self-associated multimers the 80-Å particles seen by freeze-fracture electron microscopy) which results in a significantly larger bilayer defect than do tryptic insoluble peptide monomers. The failure of complete saturation of efflux by the fast component is ascribed to the presence of two populations of small unilamellar vesicles, some of which contain tryptic insoluble peptide multimers and some of which do not.Addition of cholesterol to the tryptic insoluble peptide/phosphatidylcholine vesicles decreases the rate of ²²Na⁺ efflux by inhibiting primarily the fast component. Freeze-fracture electron microscopy indicates that the presence of cholesterol has no effect on the size, number or distribution of 80-Å intra-bilayer particles in the tryptic insoluble peptide/phosphatidylcholine vesicles. These results are consistent with a mechanism to explain the fast Na⁺ efflux component involving protein-lipid boundary perturbations.Efflux of ⁴⁵Ca²⁺ from phosphatidylcholine vesicles is also enhanced by incorporation of tryptic insoluble peptide, but only if divalent cations (Ca²⁺ or Mg²⁺) are present in the external bathing media as well as inside the sonicated vesicles. If monovalent Na⁺ only is present in the bathing media no ⁴⁵Ca²⁺ efflux is seen. Under conditions where ⁴⁵Ca²⁺ efflux is seen, both a fast and a slow component are present, although both appear lower than corresponding rate constants for ²²Na⁺ efflux. These results suggest a coordinated mechanism for ion efflux induced by tryptic insoluble peptide and, together with the ²²Na⁺ efflux studies, may have mechanistic implications for the transbilayer phospholipid exchange (flip-flop) suggesed to be induced at glycophorin/phospholipid interfaces (de Kruiff, B., van Zoelen, E.J.J. and van Deenen, L.L.M. (1978) Biochim. Biophys. Acta 509, 537–542).     

基于结构方程模型的森林生态系统空气负离子影响机制分析

空气负离子(Negative air ion, NAI)是衡量空气质量的重要指标之一,受到植被和环境的共同影响。然而,森林生态系统作为NAI产生的重要来源,森林中的植被和环境之间的相互作用以及对NAI的影响机制和贡献潜力仍难以捉摸。以暖温带森林生态系统中广泛分布的栓皮栎(Quercus variabilis BI.)为对象,基于自动观测设备长期定位观测获取了气象、土壤性质、空气洁净度以及植被光合等数据,利用皮尔逊相关系数分析和偏最小二乘结构方程模型分析了森林植被和环境要素对NAI的影响机制和贡献潜力。结果表明,环境要素和植被光合对NAI的贡献差异显著,植被光合对NAI的贡献潜力为62.65%,环境要素对NAI的贡献率为37.35%。环境要素中太阳辐射和饱和水汽压差的影响程度最大,分别为68.94%和16.55%。植被光合和PM_2.5主要通过直接效应影响NAI,而光合有效辐射、紫外辐射、土壤温湿度和饱和水汽压差主要通过间接效应影响NAI。因此,利用结构方程模型可以阐明植被光合与环境要素的变化对NAI的影响趋势,从而全面揭示了森林生态系统中植被产生NAI的作用机制以及...     

TRIM21 suppresses CHK1 activation by preferentially targeting CLASPIN for K63-linked ubiquitination

Expression of the E3 ligase TRIM21 is increased in a broad spectrum of cancers; however, the functionally relevant molecular pathway targeted by TRIM21 overexpression remains largely unknown. Here, we show that TRIM21 directly interacts with and ubiquitinates CLASPIN, a mediator for ATR-dependent CHK1 activation. TRIM21-mediated K63-linked ubiquitination of CLASPIN counteracts the K6-linked ubiquitination of CLASPIN which is essential for its interaction with TIPIN and subsequent chromatin loading. We further show that overexpression of TRIM21, but not a TRIM21 catalytically inactive mutant, compromises CHK1 activation, leading to replication fork instability and tumorigenesis. Our findings demonstrate that TRIM21 suppresses CHK1 activation by preferentially targeting CLASPIN for K63-linked ubiquitination, providing a potential target for cancer therapy.     

富营养化对白洋淀底栖-浮游耦合食物网结构和功能的影响

国际湖沼学的长期研究发现,一个完整的湖泊生态系统应包括底栖食物网和浮游食物网,而营养条件变化会显著改变浅水湖泊中底栖-浮游食物网的结构和功能。为了明晰富营养化对浅水湖泊底栖-浮游耦合食物网结构和功能的影响,以浅水草型湖泊——白洋淀为研究区,运用野外监测和ECOSIM与ECOPATH(Ew E)模型相结合方法,构建白洋淀底栖-浮游耦合食物网的概念模型,模拟1982—2011年间富营养化对白洋淀底栖路径和浮游路径的结构和功能影响:(1)野外监测的结果表明,从1999年至今白洋淀一直处于富营养化状态;(2)Ew E模型模拟结果表明1982—2006年,总生物量呈下降趋势,下降比例达66.38%;能流路径从以底栖路径为主转变为以浮游路径为主;(3)运用Pearson相关分析,结果表明:浮游植物与TN(r=0.67,P0.01)和TP(r=0.37,P0.05)呈显著正相关,而底栖藻类和大型沉水植物与TN(r=0.77,P0.01;r=0.67,P0.01)和TP(r=0.54,P0.01;r=0.36,P0.05)呈显著负相关。富营养化是白洋淀底栖初级和次级生产力向浮游初级和次级生产力转变的主要驱动力。采用科学的方法准确评估富营养化对湖泊底栖-浮游耦合食物网结构和功能的影响,可为湖泊生态系统管理提供技术和方法支持。     

云杉碳氮化学计量比对土壤水分和氮有效性的响应

以西南亚高山针叶林优势种——粗枝云杉(Picea asperata)为研究对象,探究不同土壤水分状况和氮添加下云杉碳氮化学计量比的变化及其响应过程。采用两因素(水分×氮素)随机区组实验,设置5个土壤水分梯度和3个氮添加浓度,其中土壤水分梯度分别是土壤田间持水量的40%(W1)、50%(W2)、60%(W3)、80%(W4)和100%(W5),氮添加浓度分别为0(N0)、20(N1)、40(N2)gNm^-2 a^-1。结果表明:(1)土壤水分和氮添加显著影响了云杉碳氮化学计量比(P<0.05),具体为:云杉植株和器官碳氮比在N0W4处理下最大值,随土壤水分有效性的降低而减小,随氮添加浓度的增加而降低。(2)随土壤水分有效性的降低,根和叶的碳含量显著升高(P<0.05),茎和叶的碳含量随着氮添加浓度的增加而降低。此外,土壤水分有效性的降低显著提高了根和茎的氮含量(P<0.05),各器官的氮含量随着氮添加浓度的增加而增加。在相同水分和氮添加浓度处理下表现为碳含量:叶>茎>根,氮含量:叶>根>茎。(3)云杉净光...     

辽东山区次生林木本植物空间分布

森林木本植物的空间格局有助于揭示群落结构的形成机制与潜在的生态学过程,且对林分经营具有一定指导意义。在0—50 m尺度范围内综合分析了辽东山区4 hm2温带次生林样地多度10的树种空间格局。研究发现:(1)在完全随机零模型下,大部分树种呈现聚集格局,聚集格局树种的比例随尺度增加而降低;在32 m的较大尺度下,随尺度增加,随机和规则格局成为树种分布的主要形式;(2)在异质性泊松过程零模型下,55.9%的树种呈现随机格局,其余大部分树种在10 m的尺度下呈现聚集格局,且随尺度增加,规则格局成为主要形式;(3)在完全随机零模型下,树种属性(林层、径级和多度)显著地影响种群聚集度,而在异质性泊松过程零模型下,树种属性对种群聚集度不存在显著影响。综上,生境异质性、扩散限制和树种属性部分解释了辽东山区次生林木本植物空间分布格局,相对而言,生境异质性的效应更为突出。研究结果有助于揭示次生林群落生物多样性的维持机制。     

Correction to: Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

Virologica Sinica - The corrected legend is given below.     

A colorimetric sensor array based on natural pigments for the discrimination of saccharides

A colorimetric sensor array based on natural pigments was developed to discriminate between various saccharides. Anthocyanins, pH‐sensitive natural pigments, were extracted from fruits and flowers and used as components of the sensor array. Variation in pH, due to the reaction between saccharides and boronic acids, caused obvious colour changes in the natural pigments. Only by observing the difference map with the naked eye could 11 common saccharides be divided into independent individuals. In conjunction with pattern recognition, the sensor array clearly differentiated between sugar and sugar alcohol with highly accuracy and allowed rapid quantification of different concentrations of maltitol and fructose. This sensor array for saccharides is expected to become a promising alternative tool for food monitoring. The link between anthocyanin and saccharide detection opened a new guiding direction for the application of anthocyanins in foods.