Structures of human insulin-degrading enzyme reveal a new substrate recognition mechanism

Insulin-degrading enzyme (IDE), a Zn2+-metalloprotease, is involved in the clearance of insulin and amyloid-beta (refs 1-3). Loss-of-function mutations of IDE in rodents cause glucose intolerance and cerebral accumulation of amyloid-beta, whereas enhanced IDE activity effectively reduces brain amyloid-beta (refs 4-7). Here we report structures of human IDE in complex with four substrates (insulin B chain, amyloid-beta peptide (1-40), amylin and glucagon). The amino- and carboxy-terminal domains of IDE (IDE-N and IDE-C, respectively) form an enclosed cage just large enough to encapsulate insulin. Extensive contacts between IDE-N and IDE-C keep the degradation chamber of IDE inaccessible to substrates. Repositioning of the IDE domains enables substrate access to the catalytic cavity. IDE uses size and charge distribution of the substrate-binding cavity selectively to entrap structurally diverse polypeptides. The enclosed substrate undergoes conformational changes to form beta-sheets with two discrete regions of IDE for its degradation. Consistent with this model, mutations disrupting the contacts between IDE-N and IDE-C increase IDE catalytic activity 40-fold. The molecular basis for substrate recognition and allosteric regulation of IDE could aid in designing IDE-based therapies to control cerebral amyloid-beta and blood sugar concentrations.     

Structure of the insulin receptor substrate IRS-1 defines a unique signal transduction protein.

Since the discovery of insulin nearly 70 years ago, there has been no problem more fundamental to diabetes research than understanding how insulin works at the cellular level. Insulin binds to the alpha subunit of the insulin receptor which activates the tyrosine kinase in the beta subunit, but the molecular events linking the receptor kinase to insulin-sensitive enzymes and transport processes are unknown. Our discovery that insulin stimulates tyrosine phosphorylation of a protein of relative molecular mass between 165,000 and 185,000, collectively called pp185, showed that the insulin receptor kinase has specific cellular substrates. The pp185 is a minor cytoplasmic phosphoprotein found in most cells and tissues; its phosphorylation is decreased in cells expressing mutant receptors defective in signalling. We have now cloned IRS-1, which encodes a component of the pp185 band. IRS-1 contains over ten potential tyrosine phosphorylation sites, six of which are in Tyr-Met-X-Met motifs. During insulin stimulation, the IRS-1 protein undergoes tyrosine phosphorylation and binds phosphatidylinositol 3-kinase, suggesting that IRS-1 acts as a multisite 'docking' protein to bind signal-transducing molecules containing Src-homology 2 and Src-homology-3 domains. Thus IRS-1 may link the insulin receptor kinase and enzymes regulating cellular growth and metabolism.     

组蛋白H3K36位点甲基转移酶识别组蛋白底物的分子机制研究进展

组蛋白H3第36位赖氨酸的甲基化修饰在染色质上含量丰富,与活跃转录以及DNA损伤修复等重要生理过程相关．H3K36位点可以被一甲基化、二甲基化和三甲基化3种形式修饰,目前已知的负责组蛋白H3K36三甲基化修饰的人源蛋白是SETD2,负责组蛋白H3K36二甲基化修饰的酶包含NSD1、NSD2和NSD3和ASH1L共4名成员．这些H3K36甲基转移酶都具有非常特异的H3K36位点选择性,因此,对调控体内H3K36甲基化修饰的水平和分布十分重要．此外,它们的表达异常与人类的多种疾病相关．因此,解析组蛋白H3K36甲基转移酶识别并修饰组蛋白底物的分子机制,对揭示这些酶参与的表观遗传调控机制及其在体内的生理功能都具有十分重要的意义．早期的研究使得人们对组蛋白H3K36甲基转移酶催化底物的机制有了较深入的认识,但是由于解析的修饰酶与底物复合物的结构较少,对这些酶特异识别组蛋白底物分子机制的认识尚有很多不足．近年来,随着冷冻电镜技术的应用,H3K36甲基转移酶与核小体底物的复合物结构相继取得了突破,极大地推进了人们对这些酶识别并催化组蛋白底物分子机制的认识．本文以这几个组蛋白H3K36甲基转移酶为主要目标,对其分子机制的最新进展进行介绍总结．      

群交叉积构造的一类新的Hopf群余代数

探讨了群交叉积C#σπH和群Smash余积C×πH构成半Hopf群余代数乃至Hopf群余代数的条件,这是著名的Radford双积在Hopf群余代数系统中的实现．     

Human cdc2 protein kinase is a major cell-cycle regulated tyrosine kinase substrate

cdc2 is a catalytic subunit of a protein kinase complex, called the M-phase promoting factor, that induces entry into mitosis and is universal among eukaryotes. In HeLa cells, cdc2 is shown to be the most abundant phosphotyrosine-containing protein and its phosphotyrosine content is subject to cell-cycle regulation. One site of cdc2 tyrosine phosphorylation in vivo is selectively phosphorylated by pp60c-src in vitro.     

基于新的视觉合并机制的目标识别方法

提出了一种新的特征合并机制,模拟初级视皮层中复杂细胞汇聚合并来自不同简单细胞的响应.该机制首先对简单细胞的线性响应进行归一化,然后将同频率和方向下不同相位简单细胞的响应进行能量合并,最后取局部邻域内响应的最大值作为合并后的输出,得到对输入刺激具有一定相位和平移不变性的不变特征.将其应用于目标识别,在MNIST手写数据库上的测试结果表明:基于新的合并机制的方法能取得更低的识别错误率,对目标的局部变化有更强的鲁棒性.     

Molecular basis of xeroderma pigmentosum group C DNA recognition by engineered meganucleases

Xeroderma pigmentosum is a monogenic disease characterized by hypersensitivity to ultraviolet light. The cells of xeroderma pigmentosum patients are defective in nucleotide excision repair, limiting their capacity to eliminate ultraviolet-induced DNA damage, and resulting in a strong predisposition to develop skin cancers. The use of rare cutting DNA endonucleases-such as homing endonucleases, also known as meganucleases-constitutes one possible strategy for repairing DNA lesions. Homing endonucleases have emerged as highly specific molecular scalpels that recognize and cleave DNA sites, promoting efficient homologous gene targeting through double-strand-break-induced homologous recombination. Here we describe two engineered heterodimeric derivatives of the homing endonuclease I-CreI, produced by a semi-rational approach. These two molecules-Amel3-Amel4 and Ini3-Ini4-cleave DNA from the human XPC gene (xeroderma pigmentosum group C), in vitro and in vivo. Crystal structures of the I-CreI variants complexed with intact and cleaved XPC target DNA suggest that the mechanism of DNA recognition and cleavage by the engineered homing endonucleases is similar to that of the wild-type I-CreI. Furthermore, these derivatives induced high levels of specific gene targeting in mammalian cells while displaying no obvious genotoxicity. Thus, homing endonucleases can be designed to recognize and cleave the DNA sequences of specific genes, opening up new possibilities for genome engineering and gene therapy in xeroderma pigmentosum patients whose illness can be treated ex vivo.     

基于一种新特征参数的说话人识别

针对传统的特征参数Mel频域倒谱系数MFCC难以满足语音信号的非平稳性问题,提出一种基于小波分析的新特征参数FPBW的提取方法.为了提高训练速度,采用正交高斯混和模型,将正交变换改到最大期望EM算法之前进行,从而减少训练时间.实验结果表明,新的特征参数FPBW优于特征参数MFCC,并且采用正交高斯混合模型进一步提高了识别性能和训练速度.     

Wza the translocon for E. coli capsular polysaccharides defines a new class of membrane protein

Many types of bacteria produce extracellular polysaccharides (EPSs). Some are secreted polymers and show only limited association with the cell surface, whereas others are firmly attached to the cell surface and form a discrete structural layer, the capsule, which envelopes the cell and allows the bacteria to evade or counteract the host immune system. EPSs have critical roles in bacterial colonization of surfaces, such as epithelia and medical implants; in addition some EPSs have important industrial and biomedical applications in their own right. Here we describe the 2.26 A resolution structure of the 340 kDa octamer of Wza, an integral outer membrane lipoprotein, which is essential for group 1 capsule export in Escherichia coli. The transmembrane region is a novel alpha-helical barrel. The bulk of the Wza structure is located in the periplasm and comprises three novel domains forming a large central cavity. Wza is open to the extracellular environment but closed to the periplasm. We propose a route and mechanism for translocation of the capsular polysaccharide. This work may provide insight into the export of other large polar molecules such as DNA and proteins.     

Qa-1 restricted recognition of foreign antigen by a gamma delta T-cell hybridoma

Distinct T-lymphocyte subsets recognize antigens in conjunction with different classes of major histocompatibility complex (MHC) glycoproteins using the T-cell receptor (TCR), a disulphide-linked heterodimer associated with the CD3 complex on the cell surface. In general, class I and class II MHC products provide a context for the recognition of foreign antigens by CD8+ and CD4+ T cells, respectively. This recognition seems to be largely dependent on alpha beta TCR heterodimers, whereas the function of the second gamma delta TCR, present on a minor subpopulation of cells, is still unknown. In the mouse, the existence of six cell-surface MHC class I products (K, D, L, Qa-1, Qa-2 and Tla) has been firmly established by serological, biochemical and genetic evidence. So far, only the most polymorphic of them, K, D and L ('classical' class I) have been reported as restriction elements for T-cell recognition of foreign antigens. The function of the relatively invariant Qa and Tla molecules remains unknown. We have made a T-helper cell hybridoma clone (DGT3) that recognizes synthetic copolymer poly(Glu50Tyr50) in the context of Qa-1 cell surface product, and has a CD4-CD8- phenotype. Our studies indicate that DGT3 cells express the gamma delta TCR on the cell surface, implicating its role in Qa-1-restricted antigen recognition. This is the first evidence that T cells can recognize foreign antigen in association with self Qa product, confirming that Qa molecules not only topologically, but also functionally, belong to the MHC.     

Methanotrophy below pH 1 by a new Verrucomicrobia species

Mud volcanoes, mudpots and fumaroles are remarkable geological features characterized by the emission of gas, water and/or semi-liquid mud matrices with significant methane fluxes to the atmosphere (10(-1) to 10(3) t y(-1)). Environmental conditions in these areas vary from ambient temperature and neutral pH to high temperatures and low pH. Although there are strong indications for biological methane consumption in mud volcanoes, no methanotrophic bacteria are known that would thrive in the hostile conditions of fumaroles (temperatures up to 70 degrees C and pH down to 1.8). The first step in aerobic methane oxidation is performed by a soluble or membrane-bound methane mono-oxygenase. Here we report that pmoA (encoding the beta-subunit of membrane-bound methane mono-oxygenase) clone libraries, made by using DNA extracted from the Solfatara volcano mudpot and surrounding bare soil near the fumaroles, showed clusters of novel and distant pmoA genes. After methanotrophic enrichment at 50 degrees C and pH 2.0 the most distant cluster, sharing less than 50% identity with any other described pmoA gene, was represented in the culture. Finally we isolated an acidiphilic methanotrophic bacterium Acidimethylosilex fumarolicum SolV belonging to the Planctomycetes/Verrucomicrobia/Chlamydiae superphylum, 'outside' the subphyla of the Alpha- and Gammaproteobacteria containing the established methanotrophs. This bacterium grows under oxygen limitation on methane as the sole source of energy, down to pH 0.8--far below the pH optimum of any previously described methanotroph. A. fumarolicum SolV has three different pmoA genes, with two that are very similar to sequences retrieved from the mudpot. Highly homologous environmental 16S rRNA gene sequences from Yellowstone Park show that this new type of methanotrophic bacteria may be a common inhabitant of extreme environments. This is the first time that a representative of the widely distributed Verrucomicrobia phylum, of which most members remain uncultivated, is coupled to a geochemically relevant reaction.     

一种新的降维算法PCA_LLE在图像识别中的应用

通过结合PCA与LLE两种降维方法,提出新的PCA_LLE算法,使它们优势互补.在手写体数字数据集上进行实验,先对数据集降维,再用K近邻算法对降维后的数据分类.实验结果表明融合两种算法的PCA_LLE降维方法较原来的PCA和LLE算法准确率均有了提升.而且新算法PCA_LLE对新样本的降维时间较LLE算法减少很多.在ORL人脸数据集上的实验表明,PCA_LLE算法较PCA,LLE算法准确率有所提高.     

一种新的线性判别准则在人脸识别中的应用

针对Fisher线性判别分析(LDA)在进行人脸识别这种小样本问题分类时常常遇到类内散度矩阵Sw奇异,而无法直接应用的问题,提出一种新的线性判别准则,即:定义一个新的准则函数,在对类内散度矩阵无奇异性要求的情况下,找到此准则函数最优的权向量.应用此判别准则和Fisher LDA方法分别在ORL人脸数据库上选取40个人的图像进行识别率的测试,Fisher LDA方法的识别率为0.95,而本文方法的识别率可以达到0.955,优于Fisher LDA方法.     

A new photonic bandgap cover for a patch antenna with a photonic bandgap substrate

A new photonic bandgap (PBG) cover for a patch antenna with a photonic bandgap substrate is introduced. The plane wave expansion method and the FDTD method were used to calculate such an antenna system. Numerical results for the input return loss, radiation pattern, surface wave, and the directivity of the antennas are presented. A comparison between the conventional patch antenna and the new PBG antenna is given. It is shown that the new PBG cover is very efficient for improving the radiation directivity. The physical reasons for the improvement are also given.     

模块化基质栽培——一种绿叶蔬菜种植的新方法

在分析了目前绿叶蔬菜生产概况和所面临主要问题的基础上,介绍了绿叶蔬菜模块化基质栽培的特点,并展望了近期绿叶蔬菜模块化基质栽培需要重点攻关的问题.