药用植物分子农场研究进展

植物分子农场可以利用植物生产具有药物用途的重组蛋白或者次生代谢化合物,应用广泛。随着对动植物中具有药物用途的代谢途径的深入解析,代谢途径中关键限速酶或调控蛋白的功能不断被明确,如何选择植物分子农场的底盘植物和遗传改造途径等问题,特别是如何协同提高植物制药产量与品质一直是植物分子农场体系建立中面临的关键科学问题。综述了药用的植物分子农场的最新研究进展,着重介绍了底盘植物的选择与药用植物分子农场的构建策略,以期为提高分子农场应用效果提供有力的科技支撑。     

HO-1与HCV的相关性研究

血红素加氧酶-1（hemeoxygenase-1,HO-1）在肝脏和脾脏高表达,可以被包括某些病毒感染在内的多种因素诱导表达,具有抗氧化、抗炎、抗凋亡等保护作用。丙型肝炎病毒（hepatitisCvirus,HCV）感染可以造成慢型肝炎、肝硬化和肝癌等疾病,对人类健康造成很大威胁。研究发现HO-1通过影响HCV的复制发挥其保护作用,同时HCV也可以反向调控HO-1的表达。尽管HO-1与HCV相互作用的分子机制还不明确,但H0—1与HCV感染相关性研究不断取得重大进展,将为HCV感染的治疗提供一种新方法。     

Protein–protein interactions between SWCNT/chitosan/EGF and EGF receptor: a model of drug delivery system

Epidermal growth factor (EGF) was used as the targeting ligand to enhance the specificity of a cancer drug delivery system (DDS) via its specific interaction with the EGF receptor (EGFR) that is overexpressed on the surface of some cancer cells. To investigate the intermolecular interaction and binding affinity between the EGF-conjugated DDS and the EGFR, 50 ns molecular dynamics simulations were performed on the complex of tethered EGFR and EGF linked to single-wall carbon nanotube (SWCNT) through a biopolymer chitosan wrapping the tube outer surface (EGFR·EGF-CS-SWCNT-Drug complex), and compared to the EGFR·EGF complex and free EGFR. The binding pattern of the EGF-CS-SWCNT-Drug complex to the EGFR was broadly comparable to that for EGF, but the binding affinity of the EGF-CS-SWCNT-Drug complex was predicted to be somewhat better than that for EGF alone. Additionally, the chitosan chain could prevent undesired interactions of SWCNT at the binding pocket region. Therefore, EGF connected to SWCNT via a chitosan linker is a seemingly good formulation for developing a smart DDS served as part of an alternative cancer therapy.     

DNA重排及体外分子进化

ＤＮＡ重排是目前为止最简便、最有效的体外定向进化技术,可以对单一基因、质粒、代谢途径、部分甚至整个基因组进行改造。本综述了ＤＮＡ重排的基本原理、特点、与其它体外进化技术的不同,着重介绍了其在体外分子进化上的广泛应用,并对应用前景进行了展望。     

Pathogenic mutations in the hydrophobic core of the human prion protein can promote structural instability and misfolding

Transmissible spongiform encephalopathies, or prion diseases, are caused by misfolding and aggregation of the prion protein PrP. These diseases can be hereditary in humans and four of the many disease-associated missense mutants of PrP are in the hydrophobic core: V180I, F198S, V203I and V210I. The T183A mutation is related to the hydrophobic core mutants as it is close to the hydrophobic core and known to cause instability. We used extensive molecular dynamics simulations of these five PrP mutants to compare their dynamics and conformations to those of the wild type PrP. The simulations highlight the changes that occur upon introduction of mutations and help to rationalize experimental findings. Changes can occur around the mutation site, but they can also be propagated over long distances. In particular, the F198S and T183A mutations lead to increased flexibility in parts of the structure that are normally stable, and the short β-sheet moves away from the rest of the protein. Mutations V180I, V210I and, to a lesser extent, V203I cause changes similar to those observed upon lowering the pH, which has been linked to misfolding. Early misfolding is observed in one V180I simulation. Overall, mutations in the hydrophobic core have a significant effect on the dynamics and stability of PrP, including the propensity to misfold, which helps to explain their role in the development of familial prion diseases.     

The determinants of stability in the human prion protein: insights into folding and misfolding from the analysis of the change in the stabilization energy distribution in different conditions

The dynamic evolution of the PrP(C) from its NMR-derived conformation to a beta-sheet-rich, aggregation-prone conformation is studied through all-atom, explicit solvent molecular dynamics in different temperature and pH conditions. The trajectories are analyzed by means of a recently introduced energy decomposition approach aimed at identifying the key residues for the stabilization and folding of the protein. It is shown that under native conditions the stabilization energy is concentrated in regions of the helices H1 and H3, whereas under misfolding conditions (low pH, high temperature, or mutations in selected sites) it is spread out over helix H2. Misfolding appears to be a rearrangement of the chain that disrupts most of the native secondary structure of the protein, producing some beta-rich conformations with an energy distribution similar to that of the native state.     

基于细菌群感效应人工构建分子开关

对生物体内已有的或者人工组装的生物合成途径进行优化操作涉及两个重要问题:代谢途径中关键酶的活性及蛋白表达水平。对于酶表达水平的研究,传统的做法是采用强启动子控制下的靶蛋白过量表达策略。靶蛋白的过量表达通常会导致细胞内积累大量的无活性包涵体,从而严重影响细胞的生理状态和相关生物途径的有效运转。针对这一问题,设计一种分子开关来精确调控生物合成过程中关键酶的表达水平,对于研究生物合成途径的代谢节律以及促进生物合成途径高效运转都具有重要的实用价值。基于细菌群落中普遍存在群感效应的基本原理并结合酶促催化的动力学特征,首先在大肠杆菌群落中建立信号分子高丝氨酸内酯(AHL)介导的细胞–细胞交流机制,将靶基因egfp置入到启动子PluxI的控制之下。在细胞生长过程中,产生的AHL累积到一定浓度启动靶基因表达。通过在细胞生长的不同阶段启动AHL降解酶AiiA的表达控制环境中信号分子AHL的浓度水平,从而控制靶基因egfp的转录效率,最终实现对靶蛋白EGFP表达水平的精确控制。通过检测细胞的生长状态、靶基因在mRNA水平、蛋白质水平的表达情况证明人工设计的分子开关可以便捷高效地控制靶基因表达水平,具有时空调节的严谨性。该分子开关有望广泛应用于代谢工程和合成生物学等研究领域中。