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Role of oxyR from Sinorhizobium meliloti in Regulating the Expression of Catalases

Acta Biochimica et Biophysica Sinica 2005年第6期37卷 421-428页

作者： Li EUO~(1.3) Ming-Sheng QI~(1.3) Shi-Yi YAO~2 Hai-Ping CHENG~2 Jia-Bi ZHU~1 Guan-Qiao YU~1 1 National Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, shanghai Institutes for biological sciences.chinese academy of sciences, shanghai 2.0032. china 2.biological sciences department, lehman college, the city university of new york, bronx, ny 10468. usa, 3 graduate school of the chinese academy of sciences, shanghai 2.0032. china National Laboratory of Plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200032 China graduate school of the chinese academy of sciencesshanghai 2.0032 China biological sciences department Lehman College the City University of New YorkBronx NY 10468 USA

The process of symbiotic nitrogen fixation results in the generation of reactive oxygen speciessuch as the superoxide anion（O2-）and hydrogen peroxide（H2O2）.The response of rhizobia to these toxicoxygen species is an important factor in nodulation and nitrogen *** Sinorhizobium meliloti,oneoxyR homologue and three catalase genes,katA,katB,and katC were detected by sequence *** gene is located next to and divergently from katA on the *** investigate the possible rolesof oxyR in regulating the expression of catalases at the transcriptional level in ***,an insertion mutantof this gene was *** mutant was more sensitive and less adaptive to H2O2than the wild typestrain,and total catalase/peroxidase activity was reduced approximately fourfold with the OxyR mutationrelative to *** activities of KatA and KatB and the expression of katA::lacZ and katB::lacZ promoterfusions were increased in the mutant strain compared with the parental strain grown in the absence of H2O2,indicating that katA and katB are repressed by ***,when exposed to H2O2,katA expressionwas also increased in both *** and Escherichia *** exposed to H2O2,OxyR is convertedfrom a reduced to an oxidized form in *** concluded that the reduced form of OxyR functions as arepressor of katA and katB ***,in the presence of H2O2,reduced OxyR is converted to theoxidized form of OxyR that then results in increased katA *** further showed that oxyR expressionis autoregulated via negative feedback.

关键词： Sinorhizobium meliloti OxyR catalase katA H2O2

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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

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Positive Charges on Lysine Residues of the Extrinsic 18 kDa Protein Are Important to Its Electrostatic Interaction with Spinach Photosystem Ⅱ Membranes

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Acta Biochimica et Biophysica Sinica 2005年第11期37卷 737-742页

作者： Jin-Peng GAO Zhen-Hua YONG Feng ZHANG Kang-Cheng RUAN Chun-He XU Gen-Yun CHEN Institute of Plant Physiology & Ecology.shanghai Institutes for biological sciences,chinese academy of sciences,shanghai 2.0032.china Laboratory of Proteomics,Institute of Biochemistry and Cell Biology,shanghai Institutes for biological sciences,chinese academy of sciences,shanghai 200031,china college of sciences,Zhejiang university.Hangzhou 310029,china graduate school of the chinese academy of sciences,shanghai 2.0032.china Institute of Plant Physiology & Ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200032 China graduate school of the chinese academy of sciencesshanghai 2.0032 China college of sciences Zhejiang UniversityHangzhou 310029 China Laboratory of Proteomics Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200031 China

To determine the contribution of charged amino acids to binding with the photosystem Ⅱcomplex(PSⅡ),the amino or carboxyl groups of the extrinsic 18 kDa protein were modified with N-succinimidyl propionate(NSP)or glycine methyl ester(GME)in the presence of a water-soluble carbodiimide,*** on isoelectric point shift,4-10 and 10-14 amino groups were modified in the presence of2 and 4 mM NSP,***,3-4 carboxyl groups were modified by reaction with *** of negatively charged carboxyl groups with GME did not alter the binding activity of theextrinsic 18 kDa ***,the NSP-modified 18 kDa protein,in which the positively charged aminogroups had been modified to uncharged methyl esters,failed to bind with the PSⅡ membrane in the presenceof the extrinsic 23 kDa *** defect can not be attributed to structural or conformational alterationsimposed by chemical modification,as the fluorescence and circular dichroism spectra among native,GME-and NSP-modified extrinsic 18 kDa proteins were ***,we have concluded that the positive chargesof lysyl residues in the extrinsic 18 kDa protein are important for its interaction with PSⅡ membranes in thepresence of the extrinsic 23 kDa ***,it was found that the negative charges of carboxylgroups of this protein did not participate in binding with the extrinsic 23 kDa protein associated with PSⅡmembranes.

关键词： extrinsic 18 kDa protein circular dichroism electrostatic interaction chemical modification

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Insulin-sensitizing effects of a novel α-methyl-α-phenoxylpropionate derivative in vitro

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Acta Pharmacologica Sinica 2007年第3期28卷 417-422页

作者： Hao-shu WU~(2,4) Juan-hong YU~(3,4) Ying-yin LI~2 Yu-she YANG~3 Qiao-jun HE~2 Yi-jia LOU~(2,5) Ru-yun JI~3 ~2department of Pharmacology and Toxicology,college of Pharmaceutical sciences,Zhejiang university,Hangzhou 310058,china ~3State Key Laboratory of Drug Research,shanghai Institute of Materia Medica,shanghai Institutes for biological sciences,graduate school of the chinese academy of sciences,shanghai 201203,china department of Pharmacology and Toxicology College of Pharmaceutical Sciences Zhejiang University Hangzhou China department of State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Graduate School of the Chinese Academy of Sciences Shanghai China

Aim:To examine the insulin sensitizing effects of a novel α-methyl-α-phenoxylpropionate derivative YY20 in insulin-sensitive cell ***:Theperoxisome proliferator-activated receptor γ（PPARγ）agonist bioactivities of YY20were detected by a preadipocyte differentiation ***-PCR and Westernblotting analysis were used to detect the expression of the target gene or *** effects of YY20 on insulin-mediated glucose consumption were determined inthe HepG2 human hepatocellular carcinoma ***:YY20 could enhancethe differentiation of preadipocytes to adipocytes and upregulate the gene ex-pression of PPARγ2,as well as the protein expression of insulin receptor sub-strate-1（IRS-1）,glucose transporter-4（GLUT4）,and adiponectin（ACRP30）.Theeffects on GLUT4 and ACRP30 could be reversed by the PPARγ inhibitor ***,YY20 efficiently reduced glucose consumptions in HepG2 cells after24 h culture,and the effects were related to insulin and YY20 ***:YY20,a potential insulin-sensitizing agent like rosiglitazone,couldenhance glucose consumption in HepG2 cells in a concentration-and insulin-dependent *** may improve the insulin resistance associated with type 2diabetes.

关键词： YY20 peroxisome proliferator-activated receptor y insulin sensitivity glucose consumption

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Identification and Characterization of GH Receptor and Serum GH-binding Protein in chinese Sturgeon (Acipenser sinensis)

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Acta Biochimica et Biophysica Sinica 2004年第12期36卷 811-816页

作者： Zhi-Yong LIAO1,2 and Shang-Quan ZHU2* 1college of Life sciences, Jiangxi Normal university, Nanchang 330027, china 2Key Laboratory of Proteomics Biochemistry and Cell Biology, shanghai Institutes for biological sciences, chinese academy of sciences, graduate school of the chinese academy of sciences, shanghai 200031, china college of Life sciences Jiangxi Normal UniversityNanchang 330027 China Key Laboratory of Proteomics Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China

chinese sturgeon, a kind of cartilage ganoid, has a history of over one billion years and it is called the living fossil of aquatic biology since it keeps some evolutionary trace. Here, we characterized the growth hormone receptor (GHR) and serum growth hormone binding protein (GHBP) of chinese sturgeon. It was shown that GHR was expressed in various tissues, mainly in hepatic, kidney and intestine tissues. GHR on the hepatic membrane has high and specific affinity for bream GH (brGH) and Scatchard analysis of the binding data showed a single class of high affinity binding site with an association constant Ka of 3.1×109 M–1. A specific band around 94 kD was detected by SDS-PAGE in cross-linking studies of membrane receptors. After incubation of chinese sturgeon serum with 125I-brGH, a 125I-brGH-GHBP complex was identified by Sephadex G-75, indicating that in the serum exists GHBP specially binding to brGH.

关键词： chinese sturgeon growth hormone receptor growth hormone binding protein ligand binding

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Identification and Categorization of Horizontally Transferred Genes in Prokaryotic Genomes

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Acta Biochimica et Biophysica Sinica 2005年第8期37卷 561-566页

作者： Shuo-Yong SHI~1 Xiao-Hui CAI~1 and Da-fu DING~(1,2*) Key Laboratory of Proteomics.Institute of Biochemistry and Cell Biology.shanghai Institutes for biological sciences,chinese academy of sciences,graduate school of the chinese academy of sciences.shanghai 200031,china ~2 school of Life Science & Technology.shanghai Jiaotong university.shanghai 200030.china Key Laboratory of Proteomics Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China school of Life Science & Technology Shanghai Jiaotong UniversityShanghai 200030 China

Horizontal gene transfer(HGT),a process through which genomes acquire genetic materialsfrom distantly related organisms,is believed to be one of the major forces in prokaryotic genome ***,systematic investigation is still scarce to clarify two basic issues about HGT:(1) what types ofgenes are transferred:and(2)what influence HGT events over the organization and evolution of ***-scale investigations of these two issues will advance the systematical understanding ofHGT in the context of prokaryotic genome *** investigated 82 genomes,we constructed anHGT database across broad evolutionary *** identified four function categories containing a highproportion of horizontally transferred genes:cell envelope,energy metabolism,regulatory functions,andtransport/binding *** biased function distribution indicates that HGT is not completely random;instead,it is under high selective pressure,required by function restraints in ***,wemapped the transferred genes onto the connectivity structure map of organism-specific pathways listed inKyoto Encyclopedia of Genes and Genomes(KEGG).Our results suggest that recruitment of transferredgenes into pathways is also selectively constrained because of the tuned interaction between original *** organization structures still conserve well through evolution even with the recruitment ofhorizontally transferred ***,in pathways whose organization were significantly affected byHGT events,the operon-like arrangement of transferred genes was found to be *** resultssuggest that operon plays an essential and directional role in the integration of alien genes into pathways.

关键词： genome evolution horizontal gene transfer function and pathway analysis

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Effects of (-)-stepholidine on NMDA receptors:comparison with haloperidol and clozapine

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Acta Pharmacologica Sinica 2007年第7期28卷 953-958页

作者： Wei-hua GU~2,Shen YANG~2,Wei-xing SHI~3,Xue-chu ZHEN~2,Guo-zhang JIN~(2,4)~2 department of Pharmacology,State Key Laboratory of Drug Research,shanghai Institute of Materia Medica,shanghai Institutes ofbiological sciences,graduate school of the chinese academy of sciences,chinese academy of sciences,shanghai 201203,china ~3 Neuropsychopharmacological Research Unit,department of Psychiatry,Yale university school of Medicine,new Haven,Connecticut06511,usa department of Pharmacology State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes of Biological Sciences Graduate School of the Chinese Academy of Sciences Chinese Academy of Sciences Shanghai China department of Psychiatry Neuropsychopharmacological Research Unit Yale University School of Medicine New Haven USA

Aim:To examine whether （-）-stepholidine （SPD） has a direct effect on the N-methyl-D-aspartic acid receptors （NMDAR） containing the NMDA receptor sub-units NR2A or NR2B and to compare its effect with those of haloperidol （Hal） andclozapine （Cloz）.Methods:NMDAR was transiently expressed in human embry-onic kidney 293 （HEK293） *** in intracellular calcium concentration([Ca2+]i) induced by NMDAR activation were monitored with Fura-2 ratio ***:SPD had no significant effects on either subunit of NMDARat a concentration of less than 100 μmol/*** selectively inhibited NMDARcontaining the NR2B subunit,whereas Cloz inhibited both subunits of *** both Hal and Cloz inhibited NR1a/NR2B receptor-mediated Ca2+influx,their effects were *** was more potent and had a faster peak effect ***:Both Hal and Cloz inhibit NMDAR-mediated function,whereasSPD produced only a little inhibition at a high *** on our otherstudies,the modulation of SPD on NMDAR function may be via D1receptoraction underlying an indirect mechanism.

关键词： (-)-stepholidine haloperidol clozapine NMDA receptor calcium imaging neuroleptics

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Identification and Characterization of hmr19 Gene Encoding a Multidrug Resistance Efflux Protein from Streptomyces hygroscopicus *** Strain 10-22

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Acta Biochimica et Biophysica Sinica 2004年第8期36卷 519-528页

作者： Lei QIN , Heng-An WANG1 , Zhong-Qin WU, Xiao-Feng ZHANG, Mei-Lei JIN,Zi-Xin DENG2, and Guo-Ping ZHAOshanghaiResearch Center of Biotechnology, shanghai Institute for biological sciences, chinese academy of sciences,graduate school of the chinese academy of sciences, shanghai 200233, china 1school of Agriculture and Biology, shanghai Jiaotong university, shanghai 201101, china 2school of Life Science and Biotechnology, shanghai Jiaotong university, shanghai 200030, china shanghai Research Center of Biotechnology Shanghai Institute for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200233 China school of Agriculture and Biology Shanghai Jiaotong UniversityShanghai 201101 China school of Life Science and Biotechnology Shanghai Jiaotong UniversityShanghai 200030 China

The hmr19 gene was cloned from Streptomyces hygroscopicus subsp. yingchengensis strain 10- 22, a bacterium strain producing agricultural antibiotics. Sequence similarity comparison indicates that hmr19 gene may encode a predicted protein with 14 putative transmembrane α-helical spanners, belonging to the drug:H+ antiporter-2 family of the major facilitator superfamily. The expression of hmr19 in the mycelium of strain 10-22 was detected by Western blotting analysis. Gene replacement technology was employed to construct an hmr19 disruption mutant. The growth inhibition test against different antibiotics indicated that the mutant strain was 5–20 fold more susceptible to tetracycline, vancomycin and mitomycin C than the parental wild type strain. The mutant took up tetracycline much faster and accumulated more antibiotics than the wild type strain 10-22. While with the addition of an energy uncoupler, carbonyl cyanide m- chlorophenylhydrazone, the characteristics of the accumulation of [3H]tetracycline in these two strains were almost the same. It was thus concluded that hmr19 encoded a multidrug resistance efflux protein.

关键词： multidrug resistance (MDR) gene replacement (knockout) Streptomyces Streptomyces hygroscopicus subsp. yingchengensis strain 10-22

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Identification and Characterization of a Rat Novel Gene RSEP4 Expressed Specifically in Central Nervous System

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Acta Biochimica et Biophysica Sinica 2004年第7期36卷 501-507页

作者： Xi-Dao WANG1,2, Ling-Wei KONG1, Zhi-Qin XIE1, Yu-Qiu ZHANG3, Zhi-Xin LIN2, Zhi-Qi ZHAO3,Lei YU4, and Nai-He JING1* 1 Laboratory of Molecular Cell Biology, Laboratory of Stem Cell Biology, Institute of Biochemistry and Cell Biology, shanghai Institutes for biological sciences, chinese academy of sciences, graduate school of the chinese academy of sciences, shanghai 200031, china school of Life Science and Biotechnology, 2 shanghai Jiaotong university, shanghai 200030, china Institute of Neurobiology, Fudan university, shanghai 200433, china 3 4 department of Cell Biology, Neurobiology and Anatomy, college of Medicine, university of Cincinnati, Cincinnati, Ohio, usa Laboratory of Molecular Cell Biology Laboratory of Stem Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China school of Life Science and Biotechnology Shanghai Jiaotong UniversityShanghai 200030 China Institute of Neurobiology Fudan UniversityShanghai 200433 China department of Cell Biology Neurobiology and Anatomy College of Medicine University of CincinnatiCincinnati Ohio USA

The low-abundantly expressed genes composed the majorities of the mRNAs expressed in the central nervous system (CNS), and were thought to be important for the normal brain functions. Through differential screening a low-abundance cDNA sublibrary with mRNA from neuropathic pain of chronic constriction injury (CCI) model, we have identified a novel rat gene, rat spinal-cord expression protein 4 gene (RSEP4). The total length of RSEP4 cDNA is 2006 bp, with a 501 nucleotide open reading frame (ORF) that encodes a 167 amino acid polypeptide. Northern blot revealed that RSEP4 was expressed specifically in the CNS. In situ hybridization showed that the mRNA of RSEP4 was strongly expressed in the CA1, CA2, CA3 and DG regions of hippocampus, the Purkinje cells of cerebellum, and the small sensory neurons of dorsal horn and large motor neurons of ventral horn of spinal cord. Over-expression of RSEP4-EGFP fusion protein in the human embryonic kidney 293T cells showed that RSEP4 protein was mainly localized in the cell cytoplasm. These results suggest that RSEP4 may play some roles in the CNS.

关键词： central nervous system (CNS) rat Northern blot in situ hybridization

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Purification and Characterization of Two Endo-β-1,4-glucanases from Mollusca,Ampullaria crossean

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Acta Biochimica et Biophysica Sinica 2005年第10期37卷 702-708页

作者： Yan-Hong LI~(1#) Rui GUO~(3#) Qiu-Yu YIN~1 Ming DING~1 Si-Liang ZHANG~3 Gen-Jun XU~(1,2) Fu-Kun ZHAO~(1,2*) 1 Key Laboratory of Proteomics,Institute of Biochemistry and Cell Biology,shanghai Institutes for biological sciences.graduate school of the chinese academy of sciences,chinese academy of sciences,shanghai 200031,china 2 college of Life Science,Zhejiang Sci-Tech university,Hangzhou 310018,china 3 Bioengineering Institute,East china university of Science and Technology,shanghai 200233,china Key Laboratory of Proteomics Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Graduate School of the Chinese Academy of Sciences Chinese Academy of SciencesShanghai 200031 China Bioengineering Institute East China University of Science and TechnologyShanghai 200233 China college of Life Science Zhejiang Sci-Tech UniversityHangzhou 310018 China

Two novel endo-β-1,4-glucanases,EG45 and EG27,were isolated from the gastric juice ofmollusca,Ampullaria crossean,by anion exchange,hydrophobic interaction,gel filtration and a secondround of anion exchange *** purified proteins EG45 and EG27 appeared as a single bandon sodium dodecylsulfate polyacrylamide gel electrophoresis with a molecular mass of 45 kDa and 27 kDa,*** optimum pH for CMC activity was 5.5 for EG45 and 4.4-4.8 for *** optimumtemperature range for EG27 was broad,between 50℃ and 60℃;for EG45 it was 50℃.The analysis on thestability of these two endo-β-1,4-glucanases showed that EG27 was acceptably stable at pH 3.0-11.0 evenwhen the incubation time was prolonged to 24 h at 30℃,whereas EG45 remained relatively stable at *** 85% of the activity of EG27 could be retained upon incubation at 60℃ for 24 ***,less than 10% residual activity of EG45 was detected at 50℃.Among different kinds of substrates,bothenzymes showed a high preference for carboxymethyl ***45,in particular,showed a carboxymethylcellulose hydrolytic activity of 146.5 IU/mg *** enzymes showed low activities to xylan (from oatspelt) and Sigmacell 101,and they were inactive to p-nitrophenyl-β-D-cellobioside,salicin and starch.

关键词： cellulase endo-β-1,4-glucanase Ampullaria crossean purification substrate specificity

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