The application of the high-performance ion-exchange chromatography for the analysis of bovine milk proteins

A high-performance ion-exchange chromatographic (HPIEC) method for the analysis of milk proteins is described. The technique provides an alternative to other separation methods. The resolution obtained for both the proteins of the casein group and bovine lactoserum is comparable with that of electrophoretic analysis on polyacrylamide gels and low-pressure chromatography.     

Production of pharmaceutical proteins in milk

There is every reason to expect that it will be possible within the next few years to begin to use farm animals to produce large quantities of some of the human proteins that are needed for the treatment of disease. Revolutionary new opportunities for the production of novel proteins in milk have been created by the development of methods for gene transfer. Exploitation of these opportunities depends upon selection and cloning of milk protein genes and identification of the sequences that govern tissue specific hormonally induced expression in the mammary gland. Studies with three genes, ovine -lactoglobulin, rat -casein and whey acidic protein of rat and mouse, suggest that they may all meet this requirement. Fragments of the ovine -lactoglobulin, murine whey acidic protein and rabbit -casein genes have directed production of novel proteins in the milk of transgenic mice, sheep, rabbits and pigs. The proteins were biologically active and usually co-migrated with authentic proteins. In early experiments, protein concentration was low, but our recent observations suggest that fusion genes containing genomic clones direct production of concentrations of protein that are suitable for commercial exploitation. In the longer term, two approaches may offer the potential of more reliable expression. Control elements capable of directing expression that is independent of site of insertion of the gene, but dependent on the number of copies of the gene, have been identified for a small number of genes. The availability of such elements for the milk protein genes would increase the reliability of gene expression considerably. Alternatively, targeted mutation of genes may allow the insertion of coding sequences within an existing gene so avoiding position effects.     

茶多酚分析方法研究与最新进展

本文对茶多酚的各种分离方法、检测方法,在各领域的应用以及发展趋势进行分析和评述,着重介绍茶多酚的主要成分—儿茶素的高效液相色谱和毛细管电泳分析方法以及茶多酚的生物活性在生活领域的应用。     

Improved separation at low temperature of glycoproteins by Con A-Sepharose affinity chromatography in the presence of sodium dodecyl sulfate (SDS)

Summary The Con A-Sepharose affinity chromatography of glycoproteins was even more effective at 4°C than that at room temperature (26–28°C) in the presence of sodium dodecyl sulfate (SDS). Application of this methodology to the seperation of several glycoproteins from SDS-solubilized membrane proteins in rat cerebellum, including a glycoprotein characteristics of the Purkinje cells, was sucessful.Acknowledgments. A part of this research was supported by a Grantin-Aid for Scientific Research from the Ministry of Education, Science and Culture (No. 58480150), and also by a grant from National Center of Nervous, Mental and Muscular Disorders (NCNMMD) of the Ministry of Health and Welfare (No.83-11) in Japan.     

A simplified method for isocratic HPLC analysis of polyamines

Summary A simple technique is described for the separation and analysis of polyamines in tissues and body fluids, utilizing precolumn clean-up on disposable CM-cellulose columns, followed by an automatable HPLC procedure. Complete separation and analysis takes 12–15 min per sample with sensitivity in the pmole range.Acknowledgments. Supported by USPHS HD-09713, EPA R811621010 and by Research Scientist Development Award DA-00006 to TAS.     

The structural and mechanistic basis for recycling of Rab proteins between membrane compartments

Rab proteins are members of the Ras superfamily of GTPases and are key regulators of intracellular vesicular transport. They undergo a cycle of GTPase activity, and this activity is interconnected to a cycle of reversible attachment to membranes. This cycle is mediated by geranylgeranylation of (usually) two C-terminal cysteines, which in turn is effected by Rab geranylgeranyltransferase in concert with REP (Rab escort protein). After delivery to their respective membranes, Rabs are activated by replacement of GDP by GTP, allowing interaction with a wide variety of effector molecules involved in vesicular transport, in particular with docking of transport vesicles to their specific target membranes. After completion of these events and GTP hydrolysis, Rabs are retrieved by GDI (GDP dissociation inhibitor) and delivered to their starting compartment. Here, the structural and mechanistic basis of events occurring in Rab delivery and cycling, and the differences between REP and GDI are discussed on the basis of recent advances in the field.Received 4 November 2004; received after revision 14 February 2005; accepted 31 March 2005     

Breaking biological symmetry in membrane proteins: The asymmetrical orientation of PsaC on the pseudo-C2 symmetric Photosystem I core

The elucidation of assembly pathways of multi-subunit membrane proteins is of growing interest in structural biology. In this study, we provide an analysis of the assembly of the asymmetrically oriented PsaC subunit on the pseudo C₂-symmetric Photosystem I core. Based on a comparison of the differences in the NMR solution structure of unbound PsaC with that of the X-ray crystal structure of bound PsaC, and on a detailed analysis of the PsaC binding site surrounding the F_X iron-sulfur cluster, two models can be envisioned for what are likely the last steps in the assembly of Photosystem I. Here, we dissect both models and attempt to address heretofore unrecognized issues by proposing a mechanism that includes a thermodynamic perspective. Experimental strategies to verify the models are proposed. In closing, the evolutionary aspects of the assembly process will be considered, with special reference to the structural arrangement of the PsaC binding surface. Received 22 October 2008; received after revision 17 November 2008; accepted 05 December 2008     

The use of canonical correlation analysis to identify the order of multivariate ARMA models: simulation and application

This paper is concerned with how canonical correlation can be used to identify the structure of a linear multivariate time series model. We describe briefly methods that use the canonical correlation technique and present simulation results in order to compare and evaluate the performance of these methods. The methods are also applied to a well‐known multivariate time series. Copyright © 2000 John Wiley & Sons, Ltd.     

卫星位置误差条件下基于约束Taylor级数迭代的地面目标定位理论性能分析

基于卫星平台的地球表面目标定位系统受到卫星位置误差的影响较大,为此,该文在卫星位置存在误差的条件下,系统给出一种推导约束Taylor级数迭代公式及其理论定位性能的数学分析框架.为了便于讨论,文中以时差观测量为范例,并在三种情形下分别推导各种用于地面目标辐射源定位的约束Taylor级数迭代公式及其相应的理论定位性能,并将该理论性能与三种情形下的约束Cram′er-Rao界进行定量比较,从而得到若干定量结论.文中讨论的三种情形包括:(I)没有卫星位置误差且没有校正源的情况;(II)卫星位置存在误差且没有校正源的情况;(III)卫星位置存在误差且存在校正源(位置精确已知)的情况.最后,文中设计若干基于时差的卫星定位实验场景用以验证算法推导和理论分析的有效性.     

International interdependence of business cycles in the manufacturing industry: The use of leading indicators for forecasting and analysis

This article introduces new leading indicators for fifteen industrialized countries which enable the business cycle in manufacturing to be forecast fairly reliably between 4 and 6 months ahead. These indicators are based on an improved variant of the NBER method, yielding a composite leading indicator characterized by less erratic movements and clear turning points. The indicators are used to explore the international interdependence of business cycles and to examine the degree to which this interdependence is affected by growing economic integration, as in the EC. For each of the countries studied, the various foreign economies affecting the local business climate are identified. Since the business cycles of some countries clearly lead those of others, this international interdependence can be used to further improve the predictive power of the leading indicators in the lagging countries.