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检索条件"任意字段=中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议"

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Fertility conversion related proteins in Temperature Sensitive Genic Male Sterile Line K121S of Brassica juncea

Fertility conversion related proteins in Temperature Sensiti...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者：刘岱张传利彭少丹林良斌昆明医科大学图书馆云南农业大学热带作物学院云南农业大学农科基础课程实验教学学中心云南农业大学农学与生物技术学院

The present study was conducted to study fertility conversion related proteins in Brassicajuncea and to reveal the molecular mechanism of temperature sensitive male sterility. Weused IEF / SDS-PAGE and PDQuest analysis software technology to study the proteins atthe three nuclear flower bud stage in temperature sensitive genic male sterile line(TGMS)K121S under infertile and fertile temperatures. Proteomic analysis of plants grown at differenttemperatures revealed that a total of 2377 proteins spots were detected at fertiletemperature and 2555 spots at the infertility temperature. A total of 1522 protein spotswere common in plants grown at the two different temperatures and 855 and 1033 spotswere specific to fertility and infertility temperatures respectively. Moreover, 1522 commonproteins spots were observed in both plant samples. Mass spectrometry analysis for 69 protein points showed that a majority of these proteins(54.4%) are associated with energymetabolism. The remainder was identified to be related to growth and development, defense,protein synthesis, transcription factors and signal transduction, cytoplasmic movementand cell division, protein folding, oxygen radical scavenging related proteins etc. Wepropose that the mechanisms of K121 S fertility conversion can be attributed to the abnormalsplicing of material and energy metabolism related proteins at the mRNA level causingabnormal function at infertile temperatures. These proteins include ribulose bisphosphatecarboxylase, ribulose phosphate epimerase-3. Moreover, the transcription of actin, myosinand APC cofactor genes were probably affected by the temperature sensitive BTB/POZ regulatory domain, which caused male sterility.

关键词： Brassica juncea thermosensitive male sterility Proteomics

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Study on Computer Aided Design of Decorsin and Its Mutants' structure-activity relationship

Study on Computer Aided Design of Decorsin and Its Mutants' ...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： XINGZHEN LAO 中国药科大学

Decorsin, which was originally isolated from the leech Macrobdella decora in the NorthAmerica, is an antagonist of platelet membrane glycoprotein GPIIb / III and a potent plateletaggregation inhibitor. Decorsin contains 39 residues in primary structure, and it alsohas been characterized as relatively small disulfide-rich protein because of its six cysteineresidues in sequence. The three-dimensional structure of decorsin mainly consists of twobeta sheets which are linked by three disulfide bonds(Cys7 to Cys15, Cys17 to Cys27,Cys22 to Cys38), and its Arg31-Gly32-Asp33(RGD)sequence is considered to be epitoperequired for binding, but its structural basis for this phenomenon is unknown. The developmentof bioinformatics and computer technology makes the computer-aided proteindrug design become more and more to be *** displaying and predicting the structures of proteins or their mutants with bioinformaticalsoftwares, we can calculate in molecular level the changes of their biological natures andactivities. And then with the corresponding docking between the receptor and its ligand,we can in-depth understanding of the interaction between them. These techniques havebroad application to the design of novel ligand peptides.A decorsin mutant of which alanine34 and asparagine35 residues have been substituted bytryptophan and arginine is built by computer-aided rational peptide design. The molecularsimulation results show that the conformation of decorsin mutant4 is very stable in thatthe three disulfides form the protein core and then put RGD sequence at the apex of aloop. Our docking result shows that decorsin plays a role in antiplatelet aggregation notonly by its RGD motif interaction with its integrinαIIbβ3 receptor, but also by the flankingamino acid residues of RGD motif in which Ala34 of its C-terminal segment was placed byTry shows the augment of interaction area with integrinαIIbβ3, meanwhile Asp35 of its Cterminalsegment was placed by Arg shows the direct inter

关键词： Mutation Decorsin

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The effects of scaffolds microenvironment on cell-targeting bioactivity of BMSC-affinity peptide for in vivo cartilage regeneration

The effects of scaffolds microenvironment on cell-targeting ...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Hongjie Huang Xin Zhang Xiaoqing Hu Jingxian Zhu Chunyan Zhou Yingfang Ao Institute of Sports Medicine Peking University Third Hospital Department of Biochemistrya nd Molecular Biology Peking University School of Basic Medical Sciences

Background: Affinity peptide is a kind of short chain polypeptides which play importantbiological activity, like drug/cell targeting, cell interaction or chemotaxis. Our previousstudies found that a heptapeptide(E7, EPLQLKM) has a high specific affinity to bone derived-mesenchymal stem cells(BMSCs) without any species specificity. However, the biologicalactivity of biofactor is highly dependent on the platform materials, which providesthe site for biofunctional implementation as well as affecting cellular function by creatingspecific microenvironments for cell growth and ***/Principal Findings: This study was designed to evaluate the effects of scaffoldsmicroenvironment on BMSCtargeting bioactivity of affinity peptide for cartilage regeneration.A biphasic material platform was constructed with chitosan hydrogel and a demineralizedbone matrix, which showed superior porosity, equilibrium swelling and biodegradationrate. E7 affinity peptide could be covalently coupled to this biomaterial. Comparedwith scrambled peptide containing the same amino acids as E7(but in a different order),E7 conjugated scaffolds significantly improved BMSC-targeting recruitment and homingin vitro and in vivo. In vitro culture study showed that E7 conjugated scaffolds prompteda significant up-regulated BMSC proliferation and chondrogenic differentiation thancontrol scaffolds, based on DNA quantification, cartilagous matrix production and hyaline-cartilage RNA expression. A rabbit knee cartilage defects model was used to determinethe influence of BMSC-targeting recruitment on cartilage repairing, which showed that,six months after implantation, the E7-conjaguted scaffolds significantly improved thequantity and quality of cartilage regeneration than routine surgery or control *** results indicated that affinity peptide and an appropriate microenvironment couldfacilitate cell targeting recruitment, proliferation, differentiation and subsequent tissue regenerati

关键词： scaffolds microenvironment cell-targeting affinity peptide functionalb iomaterials cartilage regeneration

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Compound SA79 Enhances Oct4 Activity and Reinforces Pluripotency through NF-γB Signaling Pathway

Compound SA79 Enhances Oct4 Activity and Reinforces Pluripot...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者：秦洪双鲍永利于春雷李玉新 National Engineering Laboratory for Druggable geneand Protein Screening Northeast Normal University Research Center of Agriculture and Medicine Gene Engineering of Ministry of Education Northeast Normal University Institute of Genetics and Cytology Northeast Normal University

Pluripotent stem cells are expected to provide revolutionary therapeutic applications in regenerativemedicine and drug discovery technologies. However, the trend toward differentiationof stem cells in vitro culture creates problems for their large scale production aswell as for their clinical applications. The maintenance of pluripotency in pluripotent cellsrelies on a complex network of transcription factors, with Oct4 playing a key role. In thisstudy, we constructed a firefly luciferase reporter driven by the Oct4 gene promoter toscreen small-molecule compounds capable of maintaining the self-renewal and pluripotencyof pluripotent stem cells. Then we identified compound SA79 could efficiently promotethe promoter activity of Oct4 gene, further results revealed that SA79 also efficientlypromoted the expression of Oct4 in both mRNA and protein levels. Moreover, SA79 couldpromote the formation of colony with high expression of pluripotent factors Oct4 andSox2 in P19 cells and umbilical cord mesenchymal stem cells(UC-MSC). Furthermore, ourresults suggested that SA79 activated NF-κB signaling via toll-like receptors pathway, andthe Oct4 expression was decreased following with blocking NF-κB signaling. Taken together,these results suggest that SA79 enhances Oct4 activity and reinforces pluripotency atleast partially through NF-κB signaling pathway.

关键词： pluripotency Oct4 NF-kB signaling pathway small-molecule compound

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Improving the thermostability of Escherichia coli phytase, appA, bymultipoint mutation

Improving the thermostability of Escherichia coli phytase, a...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Xi WANG Mingze YAO Mali SONG Aihua LIANG Institute of Biotechnology Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education

Escherichia coli phytase appA which hydrolyzes phytate has been widely applied as an important feed supplement, but the need to improve the thermal tolerance remains. In this study two strategies have been used to enhance the appA ***, eight residue substitutions(W46E, Q62 W, A73 P, K75 C, S146 E, R159 Y, N204 C, Y255D) were modified to stabilize the hydrophobic core and an increasing surface polarity of the appA. Second, two residue substitutions(Q258N, Q349N) were sequentially introduced to improve the the most ability of appA by enhancing its glycosylation level. AppA and its mutant, named appAM10,were expressed in Pichia pastoris. Results showed that the purified appAM10 had a specific activity of 3022 U/mg with an MW of approximately 55 k Da. The optimum pH was 4.5 and the optimum temperature was 65°C. Meanwhile, the Km and Vmax of appAM10 were 0.33 mM and 3147 U/mg, respectively. Compared with wild type appA, appAM10 showed an enhancement in thermal tolerance that appAM10 exhibited 17.5 % residual activity after treatment at 90 °C for 15 min(none for appA) and 7.5°C increases in the melting temperature(Tm). To understanding the mechanism of the improvement in its thermal tolerance obviously, conformational changes between appA and its mutant appAM10 had been investigated in detail by means of fluorescence spectra. Spectral results indicated that appAM10 had a tighter spatial structure and displayed larger hydrophobic exposure surface than appA. These conformational changes made the appAM10 more stable against heat treatment. This study expanded our knowledge about the mechanisms of phytase thermal tolerance.

关键词： Escherichia coli phytase thermostability glycosylation

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Reduction of AUF1-mediated follistatin mRNA decay during glucose starvation protects cells from apoptosis

Reduction of AUF1-mediated follistatin mRNA decay during glu...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Haojie Dong Lin Chen Jinghao Sheng Fan Zhang Jinfeng Su Xiangwei Gao Zhengping Xu Institute of Environmental Medicine Zhejiang University School of Medicinechina21Institute of Environmental MedicineHangzhou China Program in Molecular Cell Biology Zhejiang University School of Medicinechina21Institute of Environmental MedicineHangzhou China

Solid tumor development is frequently accompanied by energy-deficient conditions suchas glucose deprivation and ***(FST),a secretory protein originally identifiedfrom ovarian follicular fluid,has been suggested to be involved in tumor ***,whether it plays a role in cancer cell survival under energy-deprived conditionsremains *** this study,we demonstrated that glucose deprivation markedly enhancedthe expression and nucleolar localization of FST in HeLa *** studyshowed that the nuclear FST plays a protective role during cellular energy *** deprivation,the expression level of FST was up-regulated and the protein translocatedto the nucleoli in HeLa *** of FST to the nucleolus attenuated ribosomalRNA(rRNA)synthesis and ribosome biogenesis,which in turn delayed cell *** further study,we found that the induction of FST by glucose deprivation was dueto an increase in the half-life of its *** then identified an AU-rich element(ARE)inthe 3’UTR of FST mRNA that mediated its *** expression of FST was elevated afterknocking down AUF1 and reduced when AUF1 was further *** vitro RNA bindingassay and RNA pull down assays revealed that AUF1 interacted with FST mRNA directly viaits *** glucose deprivation,a majority of AUF1 shuttled from cytoplasm to nucleus,resulting in dissociation of AUF1 from FST mRNA and thus stabilization of FST ***,knockdown of AUF1 decreased whereas overexpression of AUF1increased glucosedeprivation-induced *** apoptosis promoting effect of AUF1 was eliminated inFST expressing ***,we identified a novel nucleolar function of FST,which is ofimportance in the modulation of cancer cell survival in response to glucose *** also provided evidence that AUF1 is a negative regulator of FST expression and participatesin the regulation of cell survival under glucose deprivation.

关键词： AUF1 follistatin mRNA stability glucose deprivation apoptosis

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29 Regulation of the HDM2-p53 Pathway by Ribosomal Protein L6 inResponse to Ribosomal Stress

29 Regulation of the HDM2-p53 Pathway by Ribosomal Protein L...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Dongmei Bai Xiaofeng Zheng Peking University

The HDM2-p53 pathway is crucial for monitoring p53 level and human pathologies. Therefore,identification of novel regulators involved in this regulatory loop is required for understandingthe dynamic regulation of p53 and treatment of human diseases. Here, wefound that the ribosomal protein L6 interacts with HDM2 and suppresses the E3 ubiquitinligase activity of HDM2, and subsequently attenuates HDM2-mediated p53 polyubiquitinationand degradation, which increases p53 stability and activity. The enhanced p53 activityfurther induces cell cycle arrest and cell growth inhibition. On the contrary, the level of p53 is dramatically decreased with the depletion of RPL6, indicating that RPL6 is essential forregulating p53 stabilization. We also found that RPL6 translocalizes from the nucleolus tonucleoplasm in response to ribosomal stress, which facilitates its binding with HDM2. Andthe interaction of RPL6 with HDM2 drives HDM2-mediated RPL6 polyubiquitination anddegradation. Longer treatment of ActD increases RPL6 ubiquitination and destabilizesRPL6, and thereby attenuates p53 response until the abundance of L6 subsides. RPL6 andHDM2 form an autoregulatory feedback loop to regulate p53 upon ribosomal stress. Together,our study identifies the crucial function of RPL6 in monitoring HDM2-p53 pathway,which presents the potential importance of RPL6 in human genetic diseases and cancers.

关键词： PRL6 HDM2 p53 ubiquitination cell growth

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Whole transcriptome sequencing(RNA-seq) reveals identity of two distinct kinds of hair follicles

Whole transcriptome sequencing(RNA-seq) reveals identity of ...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Rui-Bing Wu Hai-Sheng Wang Rui-Ping Hu Ji-Cheng Ye Xiu-Ling Deng Jian-YuZhang Rui Su Department of Biochemistry and molecular biology Inner Mongolia Medical University Inner Mongolia Key Laboratory ofAnimal Genetics Breeding and Reproduction College of Animal Science Inner Mongolia Agricultural University

Hair follicle of Cashmere goat(Capra hircus) is comprised of primary follicles and secondaryfollicles. The activity of primary follicle maintains a certain level after birth and the secondaryfollicle varies with the seasonal change. To comprehensively uncover the molecularbasis of these two kinds of hair follicles and the genes involved in hair follicle developmentand cashmere production, whole transcriptome sequencing was performed in goat primaryand secondary follicle. RNA-seq data demonstrated totally 617 differential expressedgenes between the primary follicle and the secondary follicle, of which 297 genes werehighly expressed in secondary follicle and 320 genes in primary follicle. Additionally, geneontology(GO) and pathway analysis showed that a large number genes responded to expressionactivation, translational initiation, positive regulation of DNA repair, and metabolicprocess were highly expression in secondary follicle and to the contrary, the genes involvedin autophagy, immune response, inflammatory response, chemotaxis and MAPKsignaling pathway were enriched in primary follicle. It specifies hair follicles identity well onthe gene expression level. To further understand the mechanisms for gene regulationbetween these two kinds of follicles, we paid attention to miRNAs and their targets whichplay important role in activation of secondary follicle. miRNA-Target-Network showed thatthe targets of these miRNAs were involved in several important biological events, such askeratin associated protein and enzymes of amino acid biosynthesis, indicating that these events are likely to play significant roles during hair production. Therefore, in this study, weused RNA-seq technology to investigate the whole transcriptome of two kinds of goat hairfollicles. The findings provided useful information about the hair follicle identities and uncovered different contributions of primary and secondary hair follicle to hair follice development and cash mere production. It faci

关键词： Cashmere goat Hair follicle transcriptome

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Novel inhibitors to HipA toxin protein can significantly reduce *** persistence

Novel inhibitors to HipA toxin protein can significantly red...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Tongqing Li Nin Yin Hongbo Liu Jianfeng Pei Luhua Lai Peking University

Drug tolerance is a sever threat to human health. Persisters are drug tolerant bacteriawithout any genotype variations. This kind of phenotype is a great challenge to infectiontreatment. Persister formation is highly related to toxin-antitoxin(TA) modules. Each pairof toxin and antitoxin is located in one operon. Free toxins can inhibit DNA replication ortranslation or cell wall synthesis, and confer to multidrug tolerance. Antitoxin can neutralizethe toxicity of toxin. On the same time, antitoxins are DNA binding proteins. They canbind to the regulation region of the operon and inhibit the transcription of the ***-antitoxin complex have a stronger binding property to the promoter. Analysis of thenetwork of the toxin-antitoxin modules1,2 indicate that inhibiting toxins may hold backthe formation of persisters and enhance the effects of antibiotics. Until now, there are noinhibitors of toxins have been published. And no compounds were discovered to regulateTA modules. HipA(high persistence gene A) was the first protein found to contribute toEscherichia coli persistence by phosphorylating the elongation factors EF-Tu3 and/or activatingppGpp signal pathway4. Overexpression of HipA increases the persister fraction by10,000 folds5. So we choose HipA toxin protein as the target to find inhibitors to *** used structure-based virtual screen for novel HipA inhibitors discovery. After atwo-step molecular docking, 190 compunds were selected for experimental tests. Thirty ofthe 190 selected compounds showed binding to HipA in the surface plasmon resonance(SPR) assay and were further tested in the persister assay. Four compounds were found tobe able to significantly reduce persisters without cell lethality. The best compound has anIC50 of 46 ± 2 μM, which exhibited the highest binding affinity to HipA, with the dissociationconstant(Kd) of 267 ± 96 nM. This compound can reduce the persister fraction byabout six fold. In the present study, we discovered that co

关键词： persistence toxin-antitoxin modules high persistence gene A(HipA)

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Regulation of MiR-486 to Gene PTEN Expression in Bovine Mammary Gland Development and Lactation

Regulation of MiR-486 to Gene PTEN Expression in Bovine Mamm...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Dan Li yingjun cui Ye Lin li Zhang xuejun Gao Qingzhang Li Chunmei Wang 东北农业大学

MicroRNAs(miRNAs) are conservative genes during evolution, and specific in expressionamong tissues and stages. Unique developmental feature during distinct stages of lactationmakes the mammary gland a main goal which discover the way to control the cyclesof cell, survival, proliferation, differentiation and death of ***, more and morestudies have showed that miRNAs play a key role in the development of normal mammarygland and lactation, and there will be more studies about mammary gland. The microarraydetected that the expression of miR-486 in different development stages mammary tissuesare obviously different. PTEN has been found as tumor-inhibiting factor and widely ***, we just obtain the tip of the iceberg knowledge have focused of the function ofmiRNA in normal mammary development,especially in bovine mammary *** this study, the function between PTEN and miR-486 was considered as a key point, andthe result shows that PTEN is modulated by miR-486 in bovine mammary gland. Therefor,we know that miR-486 is important in bovine mammary gland and *** experiment results showed that:In the microarray analysis of this study, we found that bta-miR-486 was higher in highquality lactation gland tissue than low quality tissue and pregnancy tissue, and expressionof bta-miR-486 was almost same between in low quality lactation stage and in *** in situ hybridization proved that bta-miR-486 was found primarily in mammarygland tissue rather than fat *** 3'UTR- luciferase reporter vector was constructedand the result of miR-486 about activity was detected by the Dual-Luciferase Reporter AssaySystem. The luciferase activity of reporter construction treated with miR-486 was decreased Indeed, our study confirmed that AKT, mTOR, and β-casein genes changes on account ofPTEN, and whether the PTEN level changed, it suggested that depended on *** of bta-miR-486 inhibited the expression of PTEN both in mRNA level

关键词： miR-486 cow mammary epithelial cells PTEN

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