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Effect of GNTI,a kappa opioid receptor antagonist,on MK-801-induced hyperlocomotion and stereotypy in mice

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Acta Pharmacologica Sinica 2006年第11期27卷 1401-1408页

作者： Chun-ting QI~2 Hong ZOU~2 Chen-hao ZHANG~2 Qing-lian XIE~2 Mei-lei JIN~(2,4)Lei YU~(3,4)2 research center for biotechnology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200233,china 3 Department of Genetics and center of Alcohol Studies,Rutgers,the State University of New Jersey 08854-8001,USA research center for biotechnology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China Department of Genetics and center of Alcohol Studies Rutgers the State University of New Jersey USA

Aim:To examine the effect of GNTI [5′-guanidinyl-17-(cyclopropylmethyl)-6,7-dehydro-4,5α-epoxy-3,14-dihydroxy-6,7-2′,3′-indolomorphinan],a selective antago-nist for the kappa opioid receptor,in the MK-801(dizocilpine maleate)-inducedbehavioral model of psychosis in schizophrenia as a way to explore the involve-ment of the kappa opioid receptor in modulating psychotic symptoms ***:Two doses of MK-801(0.3 mg/kg and 0.6 mg/kg)wereadministered by systemic injection in mice to induce psychosis-like behavior as arodent schizophrenia model,preceded by an injection of different doses of *** locomotion and stereotypy were measured as the behavioral endpoints forquantitative ***:GNTI inhibited MK-801-induced hyperlocomotionand *** particular,GNTI showed differential modulation of stereotypyinduced by 0.3 mg/kg vs 0.6 mg/kg ***:Antagonism of kappaopioid receptors attenuates MK-801-induced behavior,suggesting a potentialinvolvement of the kappa opioid receptor in psychosis-like symptoms *** appears to be a useful pharmacological tool to explore thekappa opioid receptor function in vivo.

关键词： GNTI kappa opioid receptor MK-801 schizophrenia model

Identification and classification of all potential hemolysin encoding genes and their products from Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai

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Acta Pharmacologica Sinica 2005年第4期26卷 453-461页

作者： Yi-xuan ZHANG~(2,3,4,5) Yan GENG(2,3,4,5) Bo BI~(2,4) Jian-yong HE~4 Chun-fu WU~4 Xiao-kui GUO~3 Guo-ping ZHAO~(2,4,6,7) 2 research center of biotechnology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200233,china 3 Department of Microbiology and Parasitology,shanghai Second Medical University,shanghai 200025,china 4 The School of Pharmaceutical Engineering,Shenyang Pharmaceutical University,Shenyang 110016,china 6 chinese National Human Genome center at shanghai,ZhangJiang High Tech Park,shanghai 201203,china research center of biotechnology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China Department of Microbiology and Parasitology Shanghai Second Medical University Shanghai China The School of Pharmaceutical Engineering Shenyang Pharmaceutical University Shenyang China chinese National Human Genome center at shanghai ZhangJiang High Tech Park Shanghai China

Aim:To identify and classify all potential hemolysin candidates of Leptospirainterrogans serogroup Icterohaemorrhagiae serovar ***:All of thepotential hemolysin encoding genes were characterized in *** geneswere cloned and expressed in Escherichia *** hemolytic activities of theexpressed proteins were assayed observing the hemolysis on sheep blood *** activities of the hemolysin candidates were measuredby thin-layer chromatography(TLC)and HPLC for *** and secretion of the hemolysins in L interrogans were studied byreverse transcription polymerase chain reaction,Western blot,and enzyme-linkedimmunosorbent *** and Conclusion:The hemolytic activities ofhemolysin candidates(LA0327,LA0378,LA1027,LA1029,LAI650,LA3050,LA3937,LA4004)from L interrogans strain Lai were *** were furtherdivided into two groups,sphingomyelinase hemolysins and non-sphingomyelinasehemolysins,based on their ability to hydrolyze *** of thesehemolysins were actually expressed in living L interrogans and some of themwere secreted into the *** study establishes an essential findcomplete basis for further studying the contribution of hemolysins to the patho-genesis of L interrogans.

关键词： hemolysin Leptospira interrogans sphingo myelinase

香樟树种子中两种Ⅱ型核糖体失活蛋白凝集素活性的研究

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生物化学与生物物理进展 2004年第7期31卷 650-654页

作者：杨海燕李荣秀刘望夷中国科学院上海生命科学研究院生物化学与细胞生物学研究所中国科学院上海生命科学研究院生物工程研究中心

从香樟树成熟的种子分离到两种新的Ⅱ型核糖体失活蛋白 :新丰毒蛋白和辛纳毒蛋白 .两者A链的分子质量虽然相差近一倍 ,但B链的分子质量相同 .Ⅱ型核糖体失活蛋白的A链是RNAN 糖苷酶 ,B链是凝集素 ,A链进入细胞表现毒性在很大程度上依赖... 详细信息

从香樟树成熟的种子分离到两种新的Ⅱ型核糖体失活蛋白 :新丰毒蛋白和辛纳毒蛋白 .两者A链的分子质量虽然相差近一倍 ,但B链的分子质量相同 .Ⅱ型核糖体失活蛋白的A链是RNAN 糖苷酶 ,B链是凝集素 ,A链进入细胞表现毒性在很大程度上依赖于B链的糖结合活性和特异性 .对辛纳毒蛋白和新丰毒蛋白B链的凝集素活性进行了测定和比较 .红细胞凝集实验显示辛纳毒蛋白和新丰毒蛋白具有相同的细胞凝集活性 ,半抗原抑制实验发现它们都属于半乳糖型核糖体失活蛋白 ,荧光光谱法显示它们的糖结合常数也相同 .

关键词：新丰毒蛋白辛纳毒蛋白凝集素荧光光谱法

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TNF receptor-associated factor-2 binding site is involved in TNFR75-dependent enhancement of TNFR55-induced cell death

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Cell research 2001年第3期11卷 217-222页

作者： LU FANG, JIA FANG, CHANG QING CHEN shanghai research center of biotechnology, shanghai institutes for biological sciences the chinese academy of sciences, shanghai 200233, china shanghai research center of biotechnology Shanghai Institutes for Biological Sciences the Chinese Academy of Sciences Shanghai China

TNF recepter-55 is the main mediator of TNF-induced apoptosis. TNF receptor-75-dependent induction or enhancement of cytotoxicity has been explained by intracellular signaling, 'ligand passing' , or induction of endogenous TNF. To study the function of human TNF receptor-75 (hTR75) and the interaction between human TNF receptor-55 (hTR55) and hTR75 in hTNFa-induced cytotoxicity, HEp-2 cells were transfected with bicistronic expression vector of hTR75 and its deletion mutants genes. hTNFa-induced cytotoxicity was determined by crystal violet colorimetric method. The expression of hTR75 and its deletion mutants in HEp-2 cells was demonstrated by RT-PCR and indirect ELISA. We found that the overexpressed hTR75 could significantly increase the susceptibility of HEp-2 cells to hTNFa which especially required TRAF2 binding site. hTR75 could not only mediate partial hTNFa-induced cytotoxicity independently but also fulfill an accessory role in enhancing or synergizing hTR55-mediated cytotoxicity

关键词： hTNFa hTNF receptors TRAF2 cytotoxcity signal transduction.

Display of aggregation-prone ligand binding domain of human PPAR gamma on surface of bacteriophage lambda

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Acta Pharmacologica Sinica 2006年第1期27卷 91-99页

作者： Bo KONG~(2,4) Wei-jun MA~(2,3,5)2 Institute of Health sciences.shanghai institutes for biological sciences,chinese academy of sciences and Medical School of shanghai Jiao Tong University.shanghai 200025.china 3 Shaughai research center of biotechnology,shanghai institutes for biological sciences,Chiese academy og sciences,shanghai 200233.china 4 Graduate School of the chinese academy of sciences Institute of Health sciences Shanghai Institutes for Biological Sciences Chinese Academy of Sciences and Medical School of Shanghai Jiao Tong University Shanghai China Graduate School of the chinese academy of sciences China shanghai research center of biotechnology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

Aim:To display the aggregation-prone ligand binding domain (LBD) of the hu-man peroxisome proliferator-activated receptor gamma (PPARγ) on the surface ofbacteriophages to establish an easy screening assay for the identification of PPARγ***:Plasmids were constructed for the expression of the PPARγLBD as a fusion to the N-terminus of the g3p protein of filamentous phage or theC-terminus of the capsid protein D (pD) of phage *** fusion proteinswere expressed in E coli and solubility characteristics were *** against the LBD as well as the pD protein were prepared for Westernblot analysis and phage capture ***:The pD-LBD fusion protein waspartially soluble,whereas the LBD-g3p fusion protein was detected only in theinsoluble *** pD-LBD fusion protein was efficiently incorporated inphage ***,the LBD was shown to be displayed on the surfaceof bacteriophage *** average,the pD-LBD fusion protein accounted for28% of the total pD protein in the lambda head ***:The hydro-phobic PPARγ LBD was expressed as a soluble form of fusion protein in E coli anddisplayed on the surface of bacteriophage lambda when it was fused to the lambdapD *** lambda pD fusion system could be used for improving the solu-bility of proteins that tend to form inclusion bodies when expressed in E *** phage particles displaying the LBD of PPARy may be of great value for theidentification of novel PPARγ ligands.

关键词： protein expression lambda phage surface display nuclear receptor peroxisome proliferator-activated receptors ligand binding domain

Recombinant scFv Antibodies against E Protein and N Protein of Severe Acute Respiratory Syndrome Virus

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Acta Biochimica et Biophysica Sinica 2004年第8期36卷 541-547页

作者： Hui LIU , Yan-Li DING, Wei HAN, Mei-Yun LIU, Rui-Yang TIAN, Sheng-Li YANG, and Yi GONGshanghai research center of biotechnology, shanghai institutes of biological sciences, chinese academy of sciences,Graduate School of the chinese academy of sciences, shanghai 200233, china shanghai research center of biotechnology Shanghai Institutes of Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of Sciences Shanghai 200233 China. huiliu@***

Three single chain antibodies (scFv) against the proteins of severe acute respiratory syndrome coronavirus (SARS-CoV) were isolated by phage display from an scFv antibody library. Bio-panning was carried out against immobilized purified envelope (E) and nucleocapsid (N) proteins of SARS-CoV. Their binding activity and specificity to E or N protein of SARS-CoV were characterized by phage-ELISA. Two of them, B10 and C20, could recognize non-overlapping epitopes of the E protein according to the two-site binding test result. Clone A17 could recognize N protein. The sequence of the epitope or overlapping epitope of scFv antibody A17 was PTDSTDNNQNGGRNGARPKQRRPQ. The affinity (equilibrium dissociation constant, Kd) of SARS-CoV E protein was 5.7×10–8 M for B10 and 8.9×10–8 M for C20. The affinity of A17 for N protein was 2.1×10–6 M. All three scFv antibodies were purified with affinity chromatography and determined by Western blot.

关键词： SARS-CoV antibody scFv phage display affinity chromatography

Anaerobic Induction of Isocitrate Lyase and Malate Synthase in Submerged Rice Seedlings Indicates the Important Metabolic Role of the Glyoxylate Cycle

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Acta Biochimica et Biophysica Sinica 2005年第6期37卷 406-414页

作者： Ying LU~(1.2) Yong-Rui WU~(1.2) Bin HAN~(1.3) 1 National center for Gene research. shanghai institutes for biological sciences. chinese academy of sciences. shanghai 200233. china. 2 Graduate School of the chinese academy of sciences. Beijing 100039.china 3 Institute of Plant Physiology and Ecology. shanghai institutes for biological sciences. chinese academy of sciences. shanghai 200032. china National center for Gene research Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200233 China Graduate School of the chinese academy of sciencesBeijing 100039 China Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200032 China

The glyoxylate cycle is a modified form of the tricarboxylic acid cycle that converts C2compounds into C4 dicarboxylic acids at plant developmental *** studying submerged rice seedlings,we revealed the activation of the glyoxylate cycle by identifying the increased transcripts of mRNAs of thegenes of isocitrate lyase(ICL)and malate synthase(MS),two characteristic enzymes of the *** blot analysis showed that ICL and MS were activated in the prolonged anaerobic *** activity assay of pyruvate decarboxylase and ICL in the submerged seedlings indicated an 8.8-fold and3.5-fold increase over that in the unsubmerged seedlings,*** activity assay of acetyl-coen-zyme A synthetase in the submerged seedlings indicated a 3-fold increase over that in the unsubmergedseedlings,which is important for initiating acetate ***,we concluded that the glyoxylatecycle was involved in acetate metabolism under anaerobic conditions.

关键词： Oryza sativa glyoxylate cycle isocitrate lyase malate synthase acetyl-coenzyme A synthetase anoxia

Sequence analysis of mRNA polyadenylation signals of rice genes

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chinese Science Bulletin 2006年第9期51卷 1069-1077页

作者： LU Ying1, 2, GAO Chenxi1,2,3 & HAN Bin1,3,41. National center for Gene research, shanghai institutes for biological sciences, chinese academy of sciences, shanghai 200233, china 2. Graduate School of the chinese academy of sciences, Beijing 100039, china 3. Institute of Plant Physiology and Ecology, shanghai institutes for biological sciences, chinese academy of sciences, shanghai 200032, china National center for Gene research Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China Graduate School of the chinese academy of sciences Beijing China Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

The formation of eukaryotic mRNAs involves the cleavage and polyadenylation of pre- mRNAs. To investigate the sequence requirement of putative polyadenylation signals (PASs), poly(A) sites and downstream elements (DUEs) in 3′-end-pro- cessing in rice, we compared expressed sequences tags (ESTs) with poly(A) extremity to full-length cDNA sequences and constructed a database of 12969 pre- mRNA sequences in ?40―+40 nt surrounding the poly(A) sites, which were from 9953 genes. The al- ternative poly(A) sites were revealed in approxi- mately 25% of mRNAs. Nearly 80% of pre-mRNAs showed stringent requirement of the YA (CA or UA) at poly (A) sites for polyadenylation. About 7.9% had the AAUAAA signals on ?40―?1 nt upstream of the poly(A) sites. Over 60% of mRNAs probably used the one- or two-base variants of AAUAAA hexamers as their PASs in 3′ fragments. The single-base variants of AAUGAA revealed the high frequency in 11.5% of 3′ fragments. The DUEs were detected in 90% of pre- mRNAs, especially more than half of the pre-mRNAs with multi-base variants of AAUAAA had the DUEs surrounding the poly(A) site. The location of DUE is also important for defining the cleavage site. Al- though most of the rice pre-mRNAs did not contain AAUAAA signal, the existence of downstream ele- ments ensured the efficiency of cleavage-polyade-

关键词：多聚腺苷酸序列分析 mRNA 水稻基因工程

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Comparative Analysis of Two-component Signal Transduction System in Two Streptomycete Genomes

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Acta Biochimica et Biophysica Sinica 2007年第5期39卷 317-325页

作者： WU WEI~(1,3#), Weihua WANG~(2#), Zhiwei CAO~(3#), Hong YU~3, Xiaojing WANG~1, Jing ZHAO~3, Hao TAN~3, Hao XU~3, Weihong JIANG~(2*), and Yixue LI~(1,3*) 1 Bioinformation center, Key Lab of Systems Biology, shanghai institutes for biological sciences, chinese academy of sciences, Graduate School of the chinese academy of sciences, shanghai 200031, china 2 Laboratory of Molecular Microbiology, Institute of Plant Physiology and Ecology, shanghai institutes for biological sciences, chinese academy of sciences, Graduate School of the chinese academy of sciences, shanghai 200032, china 3 shanghai center for Bioinformation Technology, shanghai 200235, china Bioinformation center Key Lab of Systems Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China shanghai center for Bioinformation Technologyshanghai 200235 China Laboratory of Molecular Microbiology Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200032 China

Species of the genus Streptomyces are major bacteria responsible for producing most natural antibiotics. Streptomyces coelicolor A3(2) and Streptomyces avermitilis were sequenced in 2002 and 2003, respectively. Two-component signal transduction systems (TCSs), consisting of a histidine sensor kinase (SK) and a cognate response regulator (RR), form the most common mechanism of transmembrane signal transduction in prokaryotes. TCSs in S. coelicolor A3(2) have been analyzed in detail. Here, we identify and classify the SK and RR of S. avermitilis and compare the TCSs with those of S. coelicolor A3(2) by computational approaches. Phylogenetic analysis of the cognate SK-RR pairs of the two species indicated that the cognate SK-RR pairs fall into four classes according to the distribution of their orthologs in other organisms. In addition to the cognate SK-RR pairs, some potential partners of non-cognate SK-RR were found, including those of unpaired SK and orphan RR and the cross-talk between different components in either strain. Our study provides new clues for further exploration of the molecular regulation mechanism of streptomycetes with industrial importance.

关键词： Streptomyces two-component system cross-talk phylogenetic analysis