Structure of the C-terminal domain of the pro-apoptotic protein Hrk and its interaction with model membranes

The protein harakiri (Hrk) is a pro-apoptotic BH3-only protein which belongs to the Bcl-2 family. Hrk appears associated to the mitochondrial outer membrane, apparently by a putative transmembrane domain, where it exerts its function. In this work we have identified a 27mer peptide supposed to be the putative membrane domain of the protein at the C-terminal region, and used infrared and fluorescence spectroscopies to study its secondary structure as well as to characterize its effect on the physical properties of phospholipid model membranes. The results presented here showed that the C-terminal region of Hrk adopts a predominantly α-helical structure whose proportion and destabilization capability varied depending on phospholipid composition. Moreover it was found that the orientation of the α-helical component of this C-terminal Hrk peptide was nearly perpendicular to the plane of the membrane. These results indicate that this domain is able of inserting into membranes, where it adopts a transmembrane α-helical structure as well as it considerably perturbs the physical properties of the membrane.     

Physicochemical interactions of amyloid β-peptide with lipid bilayers

The aggregation and deposition onto neuronal cells of amyloid β-peptide (Aβ) is central to the pathogenesis of Alzheimer's disease. Accumulating evidence suggests that membranes play a catalytic role in the aggregation of Aβ. This article summarizes the structures and properties of Aβ in solution and the physicochemical interaction of Aβ with lipid bilayers of various compositions. Reasons for discrepancies between results by different research groups are discussed. The importance of ganglioside clusters in the aggregation of Aβ is emphasized. Finally, a hypothetical physicochemical cascade in the pathogenesis of the disease is proposed.     

Characterization of the conformational and orientational dynamics of ganglioside GM1 in a dipalmitoylphosphatidylcholine bilayer by molecular dynamics simulations

The structure and dynamics of a single GM1 (Gal5-β1,3-GalNAc4-β1,4-(NeuAc3-α2,3)-Gal2-β1,4-Glc1-β1,1-Cer) embedded in a DPPC bilayer have been studied by MD simulations. Eleven simulations, each of 10 ns productive run, were performed with different initial conformations of GM1. Simulations of GM1-Os in water and of a DPPC bilayer were also performed to delineate the effects of the bilayer and GM1 on the conformational and orientational dynamics of each other. The conformation of the GM1 headgroup observed in the simulations is in agreement with those reported in literature; but the headgroup is restricted when embedded in the bilayer. NeuAc3 is the outermost saccharide towards the water phase. Glc1 and Gal2 prefer a parallel, and NeuAc3, GalNac4 and Gal5 prefer a perpendicular, orientation with respect to the bilayer normal. The overall characteristics of the bilayer are not affected by the presence of GM1; however, GM1 does influence the DPPC molecules in its immediate vicinity. The implications of these observations on the specific recognition and binding of GM1 embedded in a lipid bilayer by exogenous proteins as well as proteins embedded in lipids have been discussed.     

Mitochondrial import of Omi: the definitive role of the putative transmembrane region and multiple processing sites in the amino-terminal segment

The mitochondrial serine protease Omi/HtrA2 has a proapoptotic role in mammalian cells. However, neither the topology nor the processing of Omi in mitochondria is clearly understood. To determine the topology of Omi in the mitochondrial IMS, EGFP fusions were expressed with the entire N-terminal segment of full-length Omi (FL-Omi) (133-EGFP), and that without the transmembrane region (DeltaTM-EGFP) in the cells. Immunocytochemical staining and alkaline extraction experiments revealed that the TM determines the topology of Omi in the IMS and anchors the pro form into the inner membrane. As a result, the protease and the PDZ domains are exposed to the IMS. Mature Omi largely exists in the IMS as a soluble form. The processing sites of the precursor protein were examined by in vitro import experiments. The import of the processing mutants revealed importance of Arg80, Arg91, and Arg93 residues for the processing of the N-terminal segment of FL-Omi. These results suggest that the N-terminal segment of FL-Omi contains multiple processing sites processed by matrix processing proteases.     

Trophic structure of macrobenthos in the Tagus estuary and adjacent coastal shelf

The trophic structure of benthic communities in the Tagus estuary and adjacent coastal shelf was characterized according to a functional guild approach, based on sampling surveys conducted between 1987 and 2000. Macrobenthic organisms were assigned to seven distinct trophic groups (herbivorous, filter feeders, surface deposit feeders, subsurface deposit feeders, carnivores, filter feeders/detritivores, carnivores/detritivores) and the dominance of these groups was related to environmental variables using multivariate ordination techniques. Surface-deposit feeders were numerically dominant in the Tagus estuary, making up 52% of the benthic communities, while in the adjacent coastal shelf the assemblage was dominated by both surface-deposit feeders and filter feeders (37% and 33%, respectively). When biomass was considered, filter feeders and filter feeders/detritivores were the dominant groups in the estuary, while for the adjacent coastal shelf filter feeders represented 83% of the total biomass. Salinity, depth and sediment composition were the main factors structuring spatial distribution. Surface-deposit feeders were the most abundant macrobenthos of the upper estuary. Surface deposit feeders also dominated the middle and the lower estuary but the proportion of filter feeders as well as other trophic groups increased with salinity. Generally, a more even distribution of trophic structure was found at stations with high salinity. In the adjacent coastal shelf, the trophic diversity decreased with depth. The trophic structure revealed that filter feeders dominated in abundance and biomass in shallow sandy sediments (<25 m), while in deeper sandy mud and muddy habitats (>50 m to 260 m), deposit feeders and carnivores were the most important groups in abundance and biomass, respectively.     

Intensity enhancement of intraligand singlet-triplet π-π transition for the coordinated β-diketonate in Ni(II) complexes with a nitroxide radical

Room temperature and low temperature magnetic circular dichroism (MCD) in the intraligand spin-forbidden singlet-triplet π-π^∗ transition for the coordinated β-diketonate ligands were observed for the β-diketonato Ni(II) complexes with a chelated imino or nitronyl nitroxide radical, but not for the β-diketonato Ni(II) complexes without the radical ligands. This is elucidated by the borrowing mechanism from the singlet-singlet π-π^∗ transition through the hypothetical interligand β-diketonate-to-radical charge transfer (LLCT) in contrast to the case of Cr(III) complexes.     

含不同Anti-Waxy基因拷贝数的稻米直链淀粉含量分析

将含单拷贝Anti-Waxy基因的纯合植株分别作为父本或母本进行正反交,获得两组含双拷贝Anti-Waxy基因的杂交后代,分析了未转基因对照、转基因亲本及两组杂交后代糙米直链淀粉含量以揭示不同Anti-Waxy基因拷贝数对降低稻米直链淀粉含量程度的影响.结果显示,2个单拷贝转基因水稻糙米直链淀粉平均含量分别为10.72%和11.13%,比对照分别降低17.98%和14.84%;两组正交和反交杂交后代糙米直链淀粉平均含量分别为8.96%和8.23%,其平均值为8.60%,比2个转基因杂交亲本糙米直链淀粉含量分别降低19.78%和22.73%,比对照降低了34.20%.结果表明,增加转基因水稻基因组中Anti-Waxy基因拷贝数在一定程度上能够进一步降低稻米直链淀粉含量,通过将独立转化获得同品种转基因植株之间杂交可以成为获得高表达转基因植物的途径.     

干旱胁迫下胡杨茎流日变化分析——以塔里木河下游英苏断面为例

用PHYTALK植物生理生态监测系统对塔里木河下游英苏断面胡杨的茎流和相关环境因子的日变化进行了监测.结果表明:在极端干旱区塔里木河下游的胡杨茎流日变化表现多峰值,且夜晚仍保持一定流速;主枝与侧枝茎流日变化趋势相似,主枝流速高于侧枝;侧枝对环境变化响应更加灵敏,波动强烈,主枝相对侧枝,茎流变化响应迟钝且有明显滞后效应;茎流变化受太阳辐射、风速和大气温度影响明显,而其它环境因子如空气湿度、土壤湿度等影响则不大,气温与叶温表现出极佳相关性,气温与空气湿度呈显著负相关;同时,日变化中,不同时段起主导作用的因子不尽相同.     

毛柄小勾儿茶伴生群落种类组成及多样性研究

通过野外实地调查对小勾儿茶变种毛柄小勾儿茶(Berchemiella wilsoniivar. pubipetiolata)伴生群落4个分布点(浙江省临安市湍口和马啸、安徽省霍山县马家河和舒城县万佛山)的种类组成及物种多样性进行研究。结果表明:(1)种类丰富,但含多属或多种的大科、大属的优势地位不明显,极小科(仅含1属)、极小属(仅含1种)所占比例大;(2)属的北温带分布类型最多,科的世界分布类型比例大,仅次于泛热带分布类型;(3)4个分布群落乔木层、灌木层、草本层优势度均很低;灌木层Shannon-Wiener多样性指数较大,在4.13~4.49之间,乔木层和草本层较低。湍口、马家河、万佛山以灌木层Heip均匀度指数最大,马啸以乔木层均匀度指数最大;(4)马家河与万佛山之间的植物种类组成最相近,湍口与马啸也较相近;乔木层种类组成差异最大,灌木层最小。     

大鼠蓝斑组胺受体在应激导致颈动脉窦反射重调定中的作用

目的：揭示蓝斑（LC）的H1和H2受体在足底电击应激对颈动脉窦反射（CBR）重调定中的作用。方法：足底电击应激1周的SD大鼠,麻醉后孤离双侧颈动脉窦区,将不同窦内压（ISP）与其对应的平均动脉压（MAP）值进行Logistic五参数曲线拟合,求得ISP-MAP、ISP-增益（Gain）关系曲线及反射特征参数,观察Lc微量注射选择性H1或H2,受体拮抗剂氯苯吡胺（CHL）或西咪替丁（CIM）对应激状态下CBR的影响。结果：应激导致ISP-MAP关系曲线显著全面上移（P〈0.05）,ISP-Gain关系曲线中部明显下移（P〈0.05）,反射参数中闪压、饱和压、调定点和最大增益时的ISP值增大（P〈0.05）,而MAP反射变动范围及反射最大增益减小（P〈0.05）;LC内注射CHL（0.5μg/μl）或CIM（1.5μg/μl）20min内均可明显减弱应激对CBR的上述改变（P〈0.05）,CIM的减弱效应不如CHL的显著（P〈0.05）;LC注射上述相同剂量的CHL或CIM对非应激大鼠的CBR无明显影响（P〉0.05）;LC内注射CHL或CIM均不能使应激的CBR水平完全恢复到相应的非应激对照水平。结论：应激引起CBR重调定,反射敏感性下降;部分机制可能是激活中枢纽胺能系统,LC的H1和H2受体尤为H1受体在应激对CBR的重调定机制中发挥重要作用,下丘脑-LC的组胺能通路可能是应激所致CBR重调定的下行通路之一;除此之外,应激作用中尚有其他因素的参与。