Lipid-dependent gating of a voltage-gated potassium channel

Recent studies hypothesized that phospholipids stabilize two voltage-sensing arginine residues of certain voltage-gated potassium channels in activated conformations. It remains unclear how lipids directly affect these channels. Here, by examining the conformations of the KvAP in different lipids, we showed that without voltage change, the voltage-sensor domains switched from the activated to the resting state when their surrounding lipids were changed from phospholipids to nonphospholipids. Such lipid-determined conformational change was coupled to the ion-conducting pore, suggesting that parallel to voltage gating, the channel is gated by its annular lipids. Our measurements recognized that the energetic cost of lipid-dependent gating approaches that of voltage gating, but kinetically it appears much slower. Our data support that a channel and its surrounding lipids together constitute a functional unit, and natural nonphospholipids such as cholesterol should exert strong effects on voltage-gated channels. Our first observation of lipid-dependent gating may have general implications to other membrane proteins

Formation mechanism and control of flaring in forward tube spinning

Forward tube spinning (or flow forming) is usually employed to produce cylindrically tubular components to meet the increasing requirements for manufacturing high-performance and light-weight products at low cost and short lead-time. In forward tube spinning, flaring defect may easily occur at the opening end of tubes, which would deteriorate the quality of the spun tubular parts and reduce the material utilization. In addition, an additional operation is needed to trim away the flared end of the spun tabular parts. Efficient control of flaring formation is thus a non-trivial issue in forward tube spinning process and thus become one of the critical bottleneck issues to be addressed in this unique forming process. In this study, the formation mechanism of flaring was systematically studied via finite element (FE) simulation and an in-depth understanding was thus established, which forms basis for control of flaring forming in forward tube spinning. Based on the simulated material flow behaviour, it is found that flaring is formed by the material in non-spun zone flowing away from the mandrel. This material flow behaviour is caused by the pile up and the decreasing stiffness of the non-spun zone. In addition, the effects of process parameters on flaring were investigated to reduce flaring. The results show that the smaller feed rate and thickness reduction per pass can reduce the maximum flaring to a certain extent, but is very limited. To increase productivity and shorten forming lead-time, an efficient method to control flaring was proposed using a pressing ring in front of the roller based on the formation mechanism of flaring. FE simulation was further used to study the feasibility and demonstrates the validity of the method in terms of reducing and even eliminating the flaring with a short production lead-time. Finally, the forward tube spinning experiments were carried out to validate the formation mechanism of flaring and the method to avoid or eliminate the flaring formation in forward tube spinning

The Biochemistry, Ultrastructure, and Subunit Assembly Mechanism of AMPA Receptors

The AMPA-type ionotropic glutamate receptors (AMPA-Rs) are tetrameric ligand-gated ion channels that play crucial roles in synaptic transmission and plasticity. Our knowledge about the ultrastructure and subunit assembly mechanisms of intact AMPA-Rs was very limited. However, the new studies using single particle EM and X-ray crystallography are revealing important insights. For example, the tetrameric crystal structure of the GluA2cryst construct provided the atomic view of the intact receptor. In addition, the single particle EM structures of the subunit assembly intermediates revealed the conformational requirement for the dimer-to-tetramer transition during the maturation of AMPA-Rs. These new data in the field provide new models and interpretations. In the brain, the native AMPA-R complexes contain auxiliary subunits that influence subunit assembly, gating, and trafficking of the AMPA-Rs. Understanding the mechanisms of the auxiliary subunits will become increasingly important to precisely describe the function of AMPA-Rs in the brain. The AMPA-R proteomics studies continuously reveal a previously unexpected degree of molecular heterogeneity of the complex. Because the AMPA-Rs are important drug targets for treating various neurological and psychiatric diseases, it is likely that these new native complexes will require detailed mechanistic analysis in the future. The current ultrastructural data on the receptors and the receptor-expressing stable cell lines that were developed during the course of these studies are useful resources for high throughput drug screening and further drug designing. Moreover, we are getting closer to understanding the precise mechanisms of AMPA-R-mediated synaptic plasticity

Pan-cancer analysis of whole genomes

Cancer is driven by genetic change, and the advent of massively parallel sequencing has enabled systematic documentation of this variation at the whole-genome scale(1-3). Here we report the integrative analysis of 2,658 whole-cancer genomes and their matching normal tissues across 38 tumour types from the Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium of the International Cancer Genome Consortium (ICGC) and The Cancer Genome Atlas (TCGA). We describe the generation of the PCAWG resource, facilitated by international data sharing using compute clouds. On average, cancer genomes contained 4-5 driver mutations when combining coding and non-coding genomic elements; however, in around 5% of cases no drivers were identified, suggesting that cancer driver discovery is not yet complete. Chromothripsis, in which many clustered structural variants arise in a single catastrophic event, is frequently an early event in tumour evolution; in acral melanoma, for example, these events precede most somatic point mutations and affect several cancer-associated genes simultaneously. Cancers with abnormal telomere maintenance often originate from tissues with low replicative activity and show several mechanisms of preventing telomere attrition to critical levels. Common and rare germline variants affect patterns of somatic mutation, including point mutations, structural variants and somatic retrotransposition. A collection of papers from the PCAWG Consortium describes non-coding mutations that drive cancer beyond those in the TERT promoter(4); identifies new signatures of mutational processes that cause base substitutions, small insertions and deletions and structural variation(5,6); analyses timings and patterns of tumour evolution(7); describes the diverse transcriptional consequences of somatic mutation on splicing, expression levels, fusion genes and promoter activity(8,9); and evaluates a range of more-specialized features of cancer genomes(8,10-18).Peer reviewe

利用桿狀病毒表達系統表達金魚生長激素I基因

Using pSXIVVI +X3 without an initiation codon as an expressing vector, an occluded recombinant Trichoplusia ni nuclear polyhedrosis virus carrying the cDNA encoding goldfish growth hormone I (gf GHI) under the control of the Syn XIV promoter, has been constructed. Immunoblot analysis showed that the molecular weight of the expressed protein was 22 5kDa as expected, revealed that the virus mediated gfGHI was synthesized in the Sf cells, larvae and secreted into the supernatant and hemolymph