A backbone‐dependent rotamer library with high (ϕ, ψ) coverage using metadynamics simulations

Backbone‐dependent rotamer libraries are commonly used to assign the side chain dihedral angles of amino acids when modeling protein structures. Most rotamer libraries are created by curating protein crystal structure data and using various methods to extrapolate the existing data to cover all possible backbone conformations. However, these rotamer libraries may not be suitable for modeling the structures of cyclic peptides and other constrained peptides because these molecules frequently sample backbone conformations rarely seen in the crystal structures of linear proteins. To provide backbone‐dependent side chain information beyond the α‐helix, β‐sheet, and PPII regions, we used explicit‐solvent metadynamics simulations of model dipeptides to create a new rotamer library that has high coverage in the (ϕ, ψ) space. Furthermore, this approach can be applied to build high‐coverage rotamer libraries for noncanonical amino acids. The resulting Metadynamics of Dipeptides for Rotamer Distribution (MEDFORD) rotamer library predicts the side chain conformations of high‐resolution protein crystal structures with similar accuracy (~80%) to a state‐of‐the‐art rotamer library. Our ability to test the accuracy of MEDFORD at predicting the side chain dihedral angles of amino acids in noncanonical backbone conformation is restricted by the limited structural data available for cyclic peptides. For the cyclic peptide data that are currently available, MEDFORD and the state‐of‐the‐art rotamer library perform comparably. However, the two rotamer libraries indeed make different rotamer predictions in noncanonical (ϕ, ψ) regions. For noncanonical amino acids, the MEDFORD rotamer library predicts the χ ₁ values with approximately 75% accuracy.     

西安市城市化对景观格局及生态系统服务价值的影响

我国处于城市化高度发展阶段,截止2018年全国城市化率达到59.58%。城市的快速扩张,显著改变了区域景观格局,深刻影响城市生态系统的供给、调节、文化、支持等服务功能。近30年来,西安市城市用地扩张554.23 km²,基于西安市景观格局变化分析,利用同心缓冲区探讨城乡梯度内景观指数与生态系统服务价值的特征及其相互关系,揭示城市发展对城市生态系统的影响。结果表明,1980—2015年西安市各景观类型面积发生明显变化,耕地、草地面积逐渐减少,建设用地面积波动增加,主要来源于优势景观耕地的转化。景观破碎度最高的区域主要分布在距市中心10 km左右的城乡结合部,斑块密度、边缘密度、景观分割度最高,且随城市扩张,拐点距离逐渐增加。林地和耕地是提供生态系统服务的主要土地类型,西安地区总生态服务价值减少了9.56亿元,耕地减少最多(6.83亿元),沿城乡梯度,总生态系统服务价值均值呈现从市中心到农村递增的趋势,土壤保持的生态服务价值增长最快;沿时间梯度上,生态服务价值均值呈现逐年减少现象;从景观格局对生态服务价值影响分析得知,多样性指数与各类型生态系统服务价值之间存在显著的相...     

重度火烧迹地微地形对土壤微生物特性的影响——以坡度和坡向为例

通过对大兴安岭重度火烧迹地不同坡度和坡向的土壤微生物群落进行调查研究,旨在揭示重度火烧迹地过火6a后森林恢复过程土壤微生物群落的变化规律与影响因素.研究结果表明:平地土壤微生物生物量碳含量(MBC)和土壤微生物生物量碳氮比(MBC/MBN)均高于坡地,其中MBC/MBN达到差异极显著水平.平地土壤微生物的代谢活性AWCD值、对31种4类碳源(糖类、脂类、氨基酸、代谢物)的利用能力和Shannon-Winner多样性指数(H')均极显著低于坡地.西坡土壤微生物AWCD值和H'高于南坡,但AWCD和H'与土壤养分、pH值、EC无显著相关关系,说明坡向可能与土壤微生物代谢活性和多样性的关系并不密切,反映了两坡向土壤微生物群落结构的相似性.坡度由于影响了土壤养分和水分条件,进而影响了土壤微生物的生物量、群落结构、物种多样性和碳源利用能力.火烧迹地恢复初期平地土壤微生物量碳高于坡地,西坡高于南坡;恢复6a后,土壤微生物量碳的差异己不显著,但土壤微生物群落结构、物种多样性以及代谢特性仍具有显著差异,这可能与地形坡度仍然显著影响土壤水分含量的因素有关.     

黄瓜膨胀素的重组表达及活性分析

目的:提高纤维素的酶水解效率和开发高效的纤维素酶水解过程。方法:采用RT-PCR方法从黄瓜胚轴细胞中分离了膨胀素S1的cDNA,并使之与毕赤酵母表达质粒pPICZ(A连接,形成重组质粒pPICZ(A-exs1。通过电转化方法,用质粒pPICZ(A-exs1转化巴氏毕赤酵母GS115,得到重组菌株P.pastoris-exs1。在该重组菌株中,膨胀素的基因通过同源重组整合在毕赤酵母的染色体上,并处于毕赤酵母甲醇氧化酶启动子的下游。重组菌株P.pastoris-exs1在甲醇诱导下可合成并分泌膨胀素。结果:培养上清液没有纤维素酶活性,但具有破坏滤纸纤维素结晶结构的能力。培养上清液与里氏木霉纤维素酶等量混合后,可使纤维素酶的滤纸酶活力提高50%。结论:采用巴氏毕赤酵母GS115重组成功表达了黄瓜膨胀素,其表达产物可以促进纤维素酶对滤纸的水解。     

Noggin and bFGF cooperate to maintain the pluripotency of human embryonic stem cells in the absence of feeder layers

Human embryonic stem (hES) cells are typically maintained on mouse embryonic fibroblast (MEF) feeders or with MEF-conditioned medium. However, these xenosupport systems greatly limit the therapeutic applications of hES cells because of the risk of cross-transfer of animal pathogens. Here we showed that the bone morphogenetic protein antagonist noggin is critical in preventing differentiation of hES cells in culture. Furthermore, we found that the combination of noggin and basic fibroblast growth factor (bFGF) was sufficient to maintain the prolonged growth of hES cells while retaining all hES cell features. Since both noggin and bFGF are expressed in MEF, our findings suggest that they may be important factors secreted by MEF for maintaining undifferentiated pluripotent hES cells. Our data provide new insight into the mechanism how hES cell self-renewal is regulated. The newly developed feeder-free culture system will provide a more reliable alternative for future therapeutic applications of hES cells.     

耐温性L-谷氨酸发酵菌种的选育

应用基因组改组技术提高,L-谷氨酸生产菌在高温发酵条件下的谷氨酸产量。以天津短杆菌T6—13变异株SW07-1为原始亲株,分别经紫外线（UV）-硫酸二乙酯（DES）和X射线诱变,获得5株耐温性能略有提高的突变菌株。经2轮基因组改组,获得耐高温（能在44℃生长）的L-谷氨酸菌株F2-50。F2—50在38℃下,摇瓶发酵40h,发酵液中L-谷氨酸浓度比原始出发菌株提高了近41％,在41℃高温下,摇瓶发酵40h,L-谷氨酸浓度比原始出发菌株提高了近2倍。     

ROCK1 induces ERK nuclear translocation in PDGF-BB-stimulated migration of rat vascular smooth muscle cells

It has been known that Rho-associated protein kinase (ROCK) signaling regulates the migration of vascular smooth muscle cells (VSMCs). However, the isoform-specific roles of ROCK and its underlying mechanism in VSMC migration are not well understood. The current study thus aimed to investigate the roles of ROCK1/2 and their relationship to the MAPK signaling pathway in platelet-derived growth factor (PDGF)-induced rat aorta VSMC migration by manipulating ROCK gene expression. The results revealed that ROCK1 small interfering ribonucleic acid (siRNA) rather than ROCK2 siRNA decreased PDGF-BB-generated VSMC migration, and upregulation of ROCK1 expression via transfection of constructed pEGFP-C1/ROCK1 plasmid further increased the migration of PDGF-BB-treated VSMCs. In PDGF-treated VSMCs, ROCK1 siRNA did not affect the phosphorylation levels of ERK and p38 in the cytoplasm, but decreased the level of ERK phosphorylation in the nucleus. These findings demonstrate that activated ROCK1 can promote VSMC migration through facilitating phosphorylation and nuclear translocation of ERK protein.     

MiRNA介导circRNA调控肿瘤的发展

CircRNA(circular RNA)是一种具有特殊环形结构的ncRNA(non-coding RNA),并具有多种生物学功能.随着研究的深入,发现circRNA能够通过海绵吸附抑制miRNA(micro RNA)的表达,进而调控各系统肿瘤的发展.此外,一种circRNA也可参与调控一种或多种miRNA的表达,这一...