Crystallizing membrane proteins using lipidic mesophases

peer-reviewedThis paper was obtained through PEER (Publishing and the Ecology of European Research) http://www.peerproject.euA detailed protocol for crystallizing membrane proteins that makes use of lipidic mesophases is described. This has variously been referred to as the lipid cubic phase or in meso method. The method has been shown to be quite general in that it has been used to solve X-ray crystallographic structures of prokaryotic and eukaryotic proteins, proteins that are monomeric, homo- and hetero-multimeric, chromophore-containing and chromophore-free, and α-helical and β-barrel proteins. Its most recent successes are the human engineered β2-adrenergic and adenosine A2A G protein-coupled receptors. Protocols are provided for preparing and characterizing the lipidic mesophase, for reconstituting the protein into the monoolein-based mesophase, for functional assay of the protein in the mesophase, and for setting up crystallizations in manual mode. Methods for harvesting micro-crystals are also described. The time required to prepare the protein-loaded mesophase and to set up a crystallization plate manually is about one hour

Solubilization and reconstitution of influenza haemagglutinin

SIGLEAvailable from British Library Document Supply Centre- DSC:D185316 / BLDSC - British Library Document Supply CentreGBUnited Kingdo

Morphological and biochemical characterization of influenza vaccines commercially available in the United Kingdom.

Four vaccines are available in the United Kingdom against influenza virus. All are subunit vaccines, defined as either split-virion or purified surface antigen vaccine; there are two of each distinct type available. Both vaccine types are less reactogenic than whole inactivated virus, with antigenicity induced by viral surface glycoproteins. Here, each of the four vaccines has been characterized by electron microscopy and SDS-PAGE analysis, giving a unique vaccine profile. Three vaccines contain internal viral nucleoprotein which, in the presence of residual haemagglutinin, may induce an influenza A virus cross-reactive cytotoxic T-cell response and thus be of value to vaccine efficacy. Residual lipid was present in three vaccines and recent evidence suggests that pyrogenicity is correlated with the presence of viral lipid with clusters of surface glycoproteins. By a combination of electron microscopic evidence and biochemical characterization, it has been possible to resolve compositional differences, not only between vaccine type, but also between each individual currently available vaccine. Hence, there is the possibility that the morphological differences characterized here may be contributory to potential reactogenic effects subsequent to vaccination

Prenatal Effects of Antiepileptic Drugs

Antiepileptic drugs (AEDs) target ion channels and neurotransmitter systems in the brain; these same targets are responsible for regulation of processes essential for brain development. In this review, experimental findings on adverse effects of AEDs in the developing mammalian brain will be presented, including interference with physiological apoptotic cell death, cell proliferation and migration, neurogenesis, axonal arborization, synaptogenesis, and synaptic plasticity

A proposal to sequence the genome of the platypus, ornithorhynchus anatinus

© 2004 National Human Genome Research InstitutePeter D.Temple-Smith, Jennifer A. Marshall Graves, Frank Grützner, Janine Deakin, Marilyn B. Renfree, Katherine Belov, Robert Miller, Randy Jirtle, Kerstin Lindblad-Toh, Eric S. Lander and Richard K. Wilso