Demonstration by genetic suppression of interaction of GroE products with many proteins

The way in which proteins attain and maintain their final form is of fundamental importance. Recent work has focused on the role of a set of ubiquitous proteins, termed chaperonins, in the assembly of phage and multisubunit proteins. The range of chaperonin action is unknown; they could interact with most cellular polypeptides or have a limited subset of protein partners. Included in the chaperonin family is the essential heat-shock regulated Escherichia coli groEL gene product. Over-expression of the groE operon in E. coli causes enhanced assembly of heterologously expressed ribulose bisphosphate carboxylase subunits and suppresses the heat-sensitive mutant phenotype of several dnaA alleles. It has been inferred that suppression of heat-sensitive mutations is confined to dnaA alleles and that this confinement could reflect an interaction between the groE operon products and a dnaA protein aggregate at the replication origin. We now report that multiple copies of the groE operon suppress mutations in genes encoding several diverse proteins. Our data indicate a general role for the groE operon products, the GroEL and GroES proteins, in the folding-assembly pathways of many proteins.     

Interaction of Divalent Metal Ions with the Adenosine Triphosphate Measured Using Nuclear Magnetic Resonance

Theinteractionofadenosinetriphosphatewithdivalentmetalionsisimportantinbiochemicalfunctions.TheintracellularfreeMg2 canbeestimated[1]byseparatingtheα-andβ-phosphatepeaksoftheATPNMR31PspectrumandbyknowingtheMgATPdissociationconstant.TheneedfordivalentmetalionsforallenzymaticreactionsinvolvingATPiswellknown,buttheroleofthemetalionsisnotclear[2].ThisstudywasconductedtoobtaininformationconcerningchangesinthestructureofadenosinetriphosphatecausedbyinteractionwithdivalentmetalionsMg2 ,Ca2 ,…     

X…Y(X=LiF,H3N,H2O; Y=HF,LiF)复合物中锂键、氢键的理论计算

在CCSD(T)/cc-pVTZ水平下，对X…Y(X=LiF,H3N,H2O; Y=HF, LiF)复合物的9个结构进行几何构型优化和红外振动频率计算. 根据定域化分子轨道、原子自然电荷、Wiberg键级的分析表明HFLiF分子中的H-F键是共价键, 而Li-F键则为离子键而非共价键. H3N…Y (Y=HF，LiF)、H2O…Y (Y=HF，LiF)中的氢键或锂键源于静电相互作用, 并非共用电子的共价键. 结合能的计算表明: 与HF相比, LiF与X (X=LiF, H2O, NH3)的结合能更高;结合能从高到低依次为 LiF > NH3 > H2O. 红外振动频率分析表明HF与NH3、H2O形成红移氢键,即 H-F键长增加, 相应的H-F伸缩振动频率降低. H3N…LiF的Li-F键键长增加同时伸缩振动频率减少. 而LiF与H2O形成锂键后，键长增加0.016 ?，而Li-F的伸缩振动频率反而增加了2 cm-1 , 即蓝移锂键.     

Sm／ZrCl4体系促进芳基二硫醚分子中S—S键的还原断裂反应

在THF介质中温和的条件下,Sm／ZrCl4体系促进芳基二硫醚S—S键还原断裂,原位生成硫负离子,进而与酰氯反应生成硫酯;与活性卤代烃反应得到硫醚;与α,β-不饱和酯（腈）得到β-硫代酯（腈）．提供了一种新的有机硫化合物的合成方法．产物结构经红外光谱、核磁共振谱得到了证实．     

Sequence of protein disulphide isomerase and implications of its relationship to thioredoxin

The formation of disulphide bonds is essential to the structure and function of proteins. These bonds rapidly form either cotranslationally or immediately post-translationally in the lumen of the endoplasmic reticulum. Native disulphide pairing for such proteins has been achieved in vitro; however, the rates of reassembly are slow and the conditions non-physiological. To account for these observations, Anfinsen et al. proposed that a 'disulphide interchange protein' was the in vivo catalyst of disulphide bond rearrangement. Other groups discovered an activity with similar characteristics that catalysed the reductive cleavage of insulin and may be associated with insulin degradation, although this result has been disputed. The enzyme involved, protein disulphide isomerase (PDI; EC 5.3.4.1), may be the in vivo catalyst of disulphide bond formation. Here we describe the sequence of cloned rat liver PDI complementary DNA which predicts a protein with two distinct regions homologous with Escherichia coli thioredoxin, a known cofactor in oxidation-reduction reactions. Each of these regions contains the presumed active site sequence Trp-Cys-Gly-His-Cys-Lys, suggesting that PDI, similar in action to thioredoxin, catalyses disulphide bond interchange via an internal disulphide-sulphydryl interchange. The cDNA predicts a signal peptide consistent with the view that PDI is a luminal endoplasmic reticulum protein. PDI messenger RNA, although ubiquitous, is more highly concentrated in secretory cells.     

无向超环面网的距离参数(英文)

平均距离μ(G),距离控制数γl(G)和距离独立数αd(G)是度量网络性能的重要参数.n维无向超环面网是超立方体的推广.证明了μ(G)=1/d1d2…dn-1n∑i=1(ei2+ei+ei'2-ei'/2·d1d2…dn/di),γ(G)=2当且仅当[e1'+e2'…+en'/2]≤l≤d(G)-1(d1≥d2≥…dn≥4),以及αd(G)=2当[d1+d2+…+dn-2/3]≤d≤d(G)-1(d1≥d2≥…dn≥3).     

O-甲基 α-(取代苯氨基)烃基膦酸特丁基铵的合成与杀菌活性

为了研究α-(取代苯氨基)烃基膦酸酯衍生物的结构活性关系,以取代苯胺和取代苯甲醛为起始原料,经高氯酸镁催化,与亚磷酸二甲酯反应得到α-(取代苯氨基)烃基膦酸酯1,再与叔丁胺反应合成了11个未见文献报道的O-甲基 α-(取代苯氨基)烃基膦酸特丁基铵2.通过1H NMR,13C NMR,IR,MS和元素分析对所合成的化合物进行了结构表征.初步的杀菌活性测试结果表明：大多数目标化合物具有较好的杀菌活性,并且其对真菌的防治效果优于对细菌的防治效果.在500 mg/mL的剂量下,化合物2b和2f对番茄晚疫病的防效达到75%以上,略低于对照药剂烯酰吗啉.     

A Study on the Molecular Switch of Gene Expression of the Mouse Heart Nuclear DNA Fragments

It is observed by in situ stain that LDH(1-5)…nNAD+ can probably enter the nucleopore and can be bound bound specifically with the genes that encode them. During the in vitro expression, the dilution of heart nuclear DNA fragments could enhance the expression activity of LDH/DNA and the amount of expressed LDH(1-5) is in proportion to the amount of dissociable LDH(1-5) on the LDH/DNA. With the integration of 14CLeu to the proteins, it is also observed that the addition of LDH(1-5)…nNAD+ can suppress the in vitro expression activity of LDH/DNA. AFM bservation shows that the regulation sequence at the both ends of active genes may be bound with such active factors as proteins encoded by the genes which probably is the main molecular switch of gene expression and regulation we have been always searching for. Our work shows the prospective application of the combination of AFM and isotope labeling in the research of biological reaction.     

Prolyl 4-hydroxylation regulates Argonaute 2 stability

Human Argonaute (Ago) proteins are essential components of the RNA-induced silencing complexes (RISCs). Argonaute 2 (Ago2) has a P-element-induced wimpy testis (PIWI) domain, which folds like RNase H and is responsible for target RNA cleavage in RNA interference. Proteins such as Dicer, TRBP, MOV10, RHA, RCK/p54 and KIAA1093 associate with Ago proteins and participate in small RNA processing, RISC loading and localization of Ago proteins in the cytoplasmic messenger RNA processing bodies. However, mechanisms that regulate RNA interference remain obscure. Here we report physical interactions between Ago2 and the alpha-(P4H-alpha(I)) and beta-(P4H-beta) subunits of the type I collagen prolyl-4-hydroxylase (C-P4H(I)). Mass spectrometric analysis identified hydroxylation of the endogenous Ago2 at proline 700. In vitro, both Ago2 and Ago4 seem to be more efficiently hydroxylated than Ago1 and Ago3 by recombinant human C-P4H(I). Importantly, human cells depleted of P4H-alpha(I) or P4H-beta by short hairpin RNA and P4H-alpha(I) null mouse embryonic fibroblast cells showed reduced stability of Ago2 and impaired short interfering RNA programmed RISC activity. Furthermore, mutation of proline 700 to alanine also resulted in destabilization of Ago2, thus linking Ago2 P700 and hydroxylation at this residue to its stability regulation. These findings identify hydroxylation as a post-translational modification important for Ago2 stability and effective RNA interference.     

“杜西结论”的推广

研究和推广"杜西结论",设α∈φn={(a1,a2,…,an)|ai∈N,i=1,2,…,n},定义杜西变换:D(α)=(|a1-a2|,|a2-a3|,…,|an-a1|),利用离散动力系统的分析方法,研究更一般的问题,得出结论:n(n=2k,(k=1,2,…))个自然数形成一个环形,再进行相邻两数大数减小数,则在有限步内n个数必都变为零,即对任意α∈φn,当n=2k,(k=1,2,…)时,存在m∈N,有Dm(α)=θ,并得出几个相关的结论.     

一类高阶有理差分方程的全局渐近稳定性

研究差分方程xn+1=xn+αxn-k/Axn+Bxn-k,n=0,1,2,…,所有正解的局部稳定性、素二周期解、有界性、不变区间和全局渐近稳定性,其中α,A,B∈(0,∞),k∈{1,2,3,…},初始条件x-k,…,x0是任意的正整数.获得了此方程的唯一正平衡点是全局渐近稳定的.     

广义轮型完全多部图的生成树数

证明了如下结论:设KWk,n是由轮图集W={Wn1,Wn2,…,Wnk}生成的n阶广义轮型完全k-部图,其中n={n1,n2,…,nk},n=|n|=n1+n2+…+nk,1≤k≤n.那么KWk,n的生成树数目为t(KWk,n)=n2k-2∏ki=1αni-1i+βni-1i-2n-ni+1,其中αi=(di+d2i-4)/2,βi=(di-d2i-4)/2,di=n-ni+3.     

赋范空间中有限个渐近一致φ-伪压缩映象公共不动点的迭代逼近

设X是一实赋范空间,D是X的非空凸子集．Ti：D→D（i=1,2,…,m）是m个渐近一致φ-伪压缩的一致L-Lipschitzian映象．证明了在一定条件下,关于{xn}的迭代：xn＋1=（1-α1,n）xn＋α1,n T1^ny1,n;y1,n（=1-α2,n）xn＋α2,nT2^ny2,n;…;ym-1,n=（1-αm,n）xn＋αm,n Tm^xxn, n≥0强收敛于有限个渐近-致φ-伪压缩的一致L—Lipschitzian映象Ti（i=1,2,…,m）的公共不动点．     

Crystal structure of a hairpin ribozyme-inhibitor complex with implications for catalysis

The hairpin ribozyme catalyses sequence-specific cleavage of RNA. The active site of this natural RNA results from the docking of two irregular helices: stems A and B. One strand of stem A harbours the scissile bond. The 2.4 A resolution structure of a hairpin ribozyme-inhibitor complex reveals that the ribozyme aligns the 2'-OH nucleophile and the 5'-oxo leaving group by twisting apart the nucleotides that flank the scissile phosphate. The base of the nucleotide preceding the cleavage site is stacked within stem A; the next nucleotide, a conserved guanine, is extruded from stem A and accommodated by a highly complementary pocket in the minor groove of stem B. Metal ions are absent from the active site. The bases of four conserved purines are positioned potentially to serve as acid-base catalysts. This is the first structure determination of a fully assembled ribozyme active site that catalyses a phosphodiester cleavage without recourse to metal ions.     

论冀中地区O、C—P凝析气藏的形成条件

据凝析油轻烃组成、甲烷及其同系物碳同位素组成、天然气与源岩吸附气指纹时比、双环倍半萜组合特征及地质结构的综合分析,查明O、C—P大多数提析气藏的油气主要来源于石炭—二叠系煤系腐植型有机质,混入了部分下第三系的油气。对石炭—二叠系成烃特点的分析表明,R。=0.6%～1.3%,以成气为主,生油为辅,原始油气比一般大于1OOCm3/t。成熟期的石炭—二叠系有机质的成烃特点和具备油气富集的良好地质条件是O、C—P凝析气藏形成的基本条件。     

Adenylylation control by intra- or intermolecular active-site obstruction in Fic proteins

Fic proteins that are defined by the ubiquitous FIC (filamentation induced by cyclic AMP) domain are known to catalyse adenylylation (also called AMPylation); that is, the transfer of AMP onto a target protein. In mammalian cells, adenylylation of small GTPases through Fic proteins injected by pathogenic bacteria can cause collapse of the actin cytoskeleton and cell death. It is unknown how this potentially deleterious adenylylation activity is regulated in the widespread Fic proteins that are found in all domains of life and that are thought to have critical roles in intrinsic signalling processes. Here we show that FIC-domain-mediated adenylylation is controlled by a conserved mechanism of ATP-binding-site obstruction that involves an inhibitory α-helix (α(inh)) with a conserved (S/T)XXXE(G/N) motif, and that in this mechanism the invariable glutamate competes with ATP γ-phosphate binding. Consistent with this, FIC-domain-mediated growth arrest of bacteria by the VbhT toxin of Bartonella schoenbuchensis is intermolecularly repressed by the VbhA antitoxin through tight binding of its α(inh) to the FIC domain of VbhT, as shown by structure and function analysis. Furthermore, structural comparisons with other bacterial Fic proteins, such as Fic of Neisseria meningitidis and of Shewanella oneidensis, show that α(inh) frequently constitutes an amino-terminal or carboxy-terminal extension to the FIC domain, respectively, partially obstructing the ATP binding site in an intramolecular manner. After mutation of the inhibitory motif in various Fic proteins, including the human homologue FICD (also known as HYPE), adenylylation activity is considerably boosted, consistent with the anticipated relief of inhibition. Structural homology modelling of all annotated Fic proteins indicates that inhibition by α(inh) is universal and conserved through evolution, as the inhibitory motif is present in ～90% of all putatively adenylylation-active FIC domains, including examples from all domains of life and from viruses. Future studies should reveal how intrinsic or extrinsic factors modulate adenylylation activity by weakening the interaction of α(inh) with the FIC active site.     

α-synuclein蛋白与A-β肽和β-synuclein蛋白相互作用位点的预测

α-synuclein蛋白的异常聚集是引起帕金森病的重要因素.实验研究发现,α-synuclein蛋白能与多个蛋白相互作用.在相互作用的蛋白质中,有的可以促进α-synuclein的异常聚集,如A-β肽;有的可以抑制α-synuclein的异常聚集,如β-synuclein蛋白,但促进或抑制α-synuclein异常聚集的机理并不明确.利用SYBYL和AUTODOK软件模拟α-synuclein蛋白与A-β肽和β-synuclein的相互作用,找到其相互作用位点,为进一步研究α-synuclein蛋白的异常聚集提供理论依据.