Cell-free expression and in meso crystallisation of an integral membrane kinase for structure determination

Membrane proteins are key elements in cell physiology and drug targeting, but getting a high-resolution structure by crystallographic means is still enormously challenging. Novel strategies are in big demand to facilitate the structure determination process that will ultimately hasten the day when sequence information alone can provide a three-dimensional model. Cell-free or in vitro expression enables rapid access to large quantities of high-quality membrane proteins suitable for an array of applications. Despite its impressive efficiency, to date only two membrane proteins produced by the in vitro approach have yielded crystal structures. Here, we have analysed synergies of cell-free expression and crystallisation in lipid mesophases for generating an X-ray structure of the integral membrane enzyme diacylglycerol kinase to 2.28-Å resolution. The quality of cellular and cell-free-expressed kinase samples has been evaluated systematically by comparing (1) spectroscopic properties, (2) purity and oligomer formation, (3) lipid content and (4) functionality. DgkA is the first membrane enzyme crystallised based on cell-free expression. The study provides a basic standard for the crystallisation of cell-free-expressed membrane proteins and the methods detailed here should prove generally useful and contribute to accelerating the pace at which membrane protein structures are solved.     

利用模拟退火算法对200 MW供热堆堆芯核设计的优化

对 2 0 0 MW核供热堆装载模式 (燃料组件布置、可燃毒物棒根数和可燃毒物质量分数配置 )进行了优化。利用模拟退火算法和先进格林函数节块法进行多步燃耗优化计算。引入敏感性系数 ,并通过敏感性分析的方法决定优化参数 ,因此在单目标和多目标优化时均取得了明显的效果。对组件布置和可燃毒物质量分数的优化计算结果表明 ,在不改变原有的富集度和组件类型的前提下 ,与参考值相比 ,优化后的循环长度、功率峰因子和卸料燃耗均有明显的改善。该燃料管理方法不仅可用于低温堆而且也可以推广到压水堆     

也门采用溢流面板堆石坝的可行性

论述了溢流面板坝的发展概况及其特点，介绍了也门的水利工程特点，指出在也门进行溢流面板坝技术论证的必要性，以Warzan坝为实例，通过一系列数值模拟分析，论证了面板坝坝顶溢流的可行性，为也门Warzan坝的可行性设计研究提供了有实际参考价值的依据。     

Ventral neural tube cells differentiate into hepatocytes in the chick embryo

A population of ventral neural tube cells has recently been shown to migrate out of the hind brain neural tube via the vagus nerve and contribute to the developing gastrointestinal tract. Since liver is also innervated by the vagus nerve, we sought to determine if these cells also migrate into the liver. Ventral neural tube cells in the caudal hindbrain of chick embryos were tagged with a replication-deficient retroviral vector containing the LacZ gene on embryonic day 2. Embryos were processed for detection of labeled cells on embryonic day 5 and 11. Labeled cells were seen in the liver on both days and identified as hepatocytes. Previously, it was believed that all hepatocytes develop from the gut endoderm. Results of the present study show an additional source for the formation of liver cells. Received 25 August 1998; received after revision 5 November 1998; accepted 5 November 1998     

Cytochemically rich neuroglia invest metacercariae in the brain ofPhoxinus phoxinus L.

Summary Histochemical determination of neuroglia investing parasites in naturally infected fish depends upon recognition of the highly soluble or diffusible nature of the cytoplasm.Acknowledgment. M.R.L. Johnston advised me during the course of this work.     

Mutations in SEC63 cause autosomal dominant polycystic liver disease

Mutations in PRKCSH, encoding the beta-subunit of glucosidase II, an N-linked glycan-processing enzyme in the endoplasmic reticulum (ER), cause autosomal dominant polycystic liver disease. We found that mutations in SEC63, encoding a component of the protein translocation machinery in the ER, also cause this disease. These findings are suggestive of a role for cotranslational protein-processing pathways in maintaining epithelial luminal structure and implicate noncilial ER proteins in human polycystic disease.     

Microbial control of diamondback moth, Plutella xylostella L. （Lepidoptera： Yponomeutidae） using bacteria （Xenorhabdus nematophila） and its metabolites from the entomopathogenic nematode Steinernema carpocapsae

Cells and cell-free solutions of the culture filtrate of the bacterial symbiont, Xenorhabdus nematophila taken from the entomopathogenic nematode Steinernema carpocapsae in aqueous broth suspensions were lethal to larvae of the diamondback moth Plutella xylostella. Their application on leaves of Chinese cabbage indicated that the cells can penetrate into the insects in the absence of the nematode vector. Cell-free solutions containing metabolites were also proved as effective as bacterial cells suspension. The application of aqueous suspensions of cells of X.nematophila or solutions containing its toxic metabolites to the leaves represents a possible new strategy for controlling insect pests on foliage.     

Thiamine derivatives bind messenger RNAs directly to regulate bacterial gene expression

Although proteins fulfil most of the requirements that biology has for structural and functional components such as enzymes and receptors, RNA can also serve in these capacities. For example, RNA has sufficient structural plasticity to form ribozyme and receptor elements that exhibit considerable enzymatic power and binding specificity. Moreover, these activities can be combined to create allosteric ribozymes that are modulated by effector molecules. It has also been proposed that certain messenger RNAs might use allosteric mechanisms to mediate regulatory responses depending on specific metabolites. We report here that mRNAs encoding enzymes involved in thiamine (vitamin B(1)) biosynthesis in Escherichia coli can bind thiamine or its pyrophosphate derivative without the need for protein cofactors. The mRNA-effector complex adopts a distinct structure that sequesters the ribosome-binding site and leads to a reduction in gene expression. This metabolite-sensing regulatory system provides an example of a 'riboswitch' whose evolutionary origin might pre-date the emergence of proteins.     

Construction of some hypergroups from combinatorial structures

Jajcay‘s studies( 1993 ; 1994) on the automorphism groups of Cayley maps yielded a new product of groups, which he called, rotary product. Using this product, we define a hyperoperation ⊙ on the group Syme (G) , the stabilizer of the identity e ∈ G in the group Sym (G) . We prove that ( Syme (G) , ⊙) is a hypergroup and characterize the subhypergroups of this hypergroup.Finally, we show that the set of all subhypergroups of Syme ( G ) constitute a lattice under ordinary join and meet and that the minimal elements of order two of this lattice is a subgroup of Aut (G) .