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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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NMR Structure of human CTCF protein and its interaction with human Protocadherin-γ gene

NMR Structure of human CTCF protein and its interaction with...

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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： DIFEI XU 中国科学技术大学

Recent studies of genome-wide chromatin interactions have revealed that the human genomefunctions depend on its 3D ***,an 11-zinc finger tandem protein,plays a vital role in chromatin structure and *** is universal and conserved expressedin cells and plays diverse important roles in transcriptional regulation,gene imprinting,insulation,different types of gene interactions,alternative splicing etc..The protocadheringene cluster(Pcdhα,-β,and-γ)can express in a combinational way to generate extraordinarysingle-cell diversity in the vertebrate nervous *** recognizing and bindingto DNA sequence specificly,CTCF can realize a‘long-range interaction’of humanPcdhαgene via DNA looping,and thus conduct different promoter-enhancer interactionfor different transcription ***,we use solution NMR and SAXS to get somestructure information of CTCF,and further experiments are ready to get a more detailedstructure of ***,we confirmed the binding affinity of hCTCF-hPcdhαDNA in vitro,providing us the opportunity to obtain the structure of CTCF-PcdhαDNA *** that CTCF may bind to PcdhαDNA in a distinctive way to execute important functions.

关键词： CTCF NMR Structure DNA looping

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The role of PP2ACγ in Neurogenesis

The role of PP2ACγ in Neurogenesis

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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：林琳刘春英佟伟民 Chinese Academy of Medical Sciences (CAMS)&Peking Union Medical College (PUMC)

Protein phosphatase 2A(PP2A), as a heterotrimeric holoenzyme, is abundant in eukaryoticcells and involved in many processes, including cell growth, differentiation, apoptosis, cellmotility, the DNA damage response, and cell cycle progression. Although recent finding shave provided an intriguing insight into the spatial regulation of protein phosphorylation and how this can be finely orchestrated by specific kinases and phosphatases, its role in the emergence and maturation of the mammalian central nervous system(CNS), especially in neurons, remains unknown. Here, we inactivated PP2ACα specifically in the CNS of mice and found that its loss caused perinatal lethality with abnormally decreased proliferation and increased apoptosis in cortical neurons. Through morphologic and genetic approaches,we show that the enzymatic activity of PP2ACα is required for the endogenous DNA damage response during cortical development, uncovering PP2ACα-mediated DNA damage response plays an important role in neurogenesis and provides an insight into the molecular pathogenesis underlying microcephaly.

关键词： Phosphatases Cortical Neurons DNA Damage Response

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The Calcineurin B subunit(CnB) synergizes with interferon-γ toactivate RAW264.7 macrophages via integrin αM–p38 MAPK–STAT1 signaling pathway

The Calcineurin B subunit(CnB) synergizes with interferon-γ...

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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： Zhenyi Su Lan Xu Wenlong Zhang Rui Yang Yanxia Yin Youxiu Zhong Juren Chen Qun Wei Department of Biochemistry and Molecular Biology Beijing Normal University Gene engineering and Biotechnology Beijing Key Laboratory

We have shown previously that the calcineurin B subunit(CnB) plays an important role ininnate immunity, but little is known of the interactions between CnB and key ***, we report that CnB and interferon-γ(IFN-γ) have a remarkable synergistic effect oninduction of typical IFN-γ response genes(e.g., CXCL9, CXCL10, iNOS, TRAIL) in RAW264.7macrophages. We also reveal that STAT1 is a key factor orchestrating the synergy of CnBand IFN-γ, which is coactivated by CnB-engaged integrin αM-p38 MAPK signaling andIFN-γ-initiated signaling. In addition, combination of CnB and IFN-γ greatly increases thecytotoxicity of RAW264.7 macrophages.

关键词： calcineurin B subunit interferon-γ synergy STAT1 p38 MAPK integrin αM

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MeRIP-PF: An Easy-to-use Pipeline for High-resolution Peak finding in MeRIP-Seq Data

MeRIP-PF: An Easy-to-use Pipeline for High-resolution Peak f...

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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：宋述慧 Beijing Institute of Genomics CAS

Background: RNA modifications, especially methylation of the N6 position of adenosine(A)-- m6 A, represent an emerging research frontier in RNA biology. With the rapid developmentof high-throughput sequencing technology, in-depth study of m6 A distributionand function relevance becomes ***: Here, we present a high-efficiency and user-friendly analysis pipeline calledMeRIP-PF for the signal identification of MeRIP-Seq data in reference to controls. MeRIPPFprovides a statistical P-value for each identified m6 A region based on the difference ofread distribution when compared to the controls and also calculates false discovery rate(FDR) as a cut off to differentiate reliable m6 A regions from the background. Furthermore,MeRIP-PF also achieves gene annotation of m6 A signals or peaks and produce outputs inboth XLS and graphical format, which are useful for further study. MeRIP-PF is implementedin Perl and is freely available at http://***/***.

关键词： RNA modifications m6A

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A light switch to control circadian rhythms

A light switch to control circadian rhythms

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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： Liting Liting Yang Sen Liu Institute of Molecular Biology China Three Gorges University

Biological clock is an endogenous rhythmicity keeping mechanisms,which is ubiquitousphenomenon in biological *** rhythms regulate various metabolic and behavioralactivities and have evolved as an adaptation to daily changes in environmental *** is the model organism to study the molecular mechanisms of circadianand its circadian clock can be reconstituted in vitro by mixing recombinant KaiA,KaiB andKaiC proteins with ATP,producing KaiC phosphorylation and dephosphorylation cyclesthat have a regular rhythm with a 24-h *** KaiA enhances the phosphorylationactivity of KaiC while protein KaiB antagonizes this *** C-terminal KaiA domain alone is sufficient in vitro to promote the phosphorylation ofKaiC,but it also remains to be seen whether it affects the circadian clock.C-terminal domainof KaiA adopts a novel four a-helical homodimeric *** first Ja helix do notinteract with kaiC directly suggested by the crystal structure and modeling *** use Jahelix of the AsLOV2 domain to replace the first Ja helix while maintaining AsLOV2 structurein the dark but allowing kaiA to bind to kaiC proteins when the Ja helix unfolds in *** hope this light switch can control KaiC de/phosphorylation,which in turn can affectthe rhythm of this molecular oscillator.

关键词： Circadian Clock Protein Design KaiA/KaiB/KaiC

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Structural Insights into Amphioxus Immunity-related Protein BbPGRP1

Structural Insights into Amphioxus Immunity-related Protein ...

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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： Wen-Jie Wang Wang Cheng Ming Luo Hong-Mei Yu Yuxing Chen Cong-Zhao Zhou USTC

Peptidoglycan recognition proteins(PGRPs)are pattern recognition molecules which areinvolved in antimicrobial defense in most animals,including insects,echinoderms,molluscsand *** date,a number of PGRPs in insects and mammalian have been studiedin structure and *** to the previously reported PGRPs,the PGRP1fromBranchiostoma belcheri tsingtauense(BbPGRP1)are fused with a N-terminal chitin ***,we reported its 2.8 ? crystal structure of the C-terminal PGRP domain,revealinga classic overall scaffold:a centralβ-sheet composed of fiveβ-strands and threeα-***1 exhibits in-vitro hydrolytic activity towards both Lys-type and ***,BbPGRP1 can only bind to and bacteriostatic against Grampositivebacteria rather than Gram-negative ***,the binding assays enabledus to characterize two key residues in the N-terminal ChtBD1 domain,Trp53 andTrp55,involved in the recognition to ***,these data indicated that BbPGRP1is a bifunctional protein,with the N-and C-terminal domains function independently.

关键词： Amphioxus Peptidoglycan recognition protein crystal structure amidase activity Chitin-binding domain

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High-level production of biologically active human enterokinase light chain by Esherichia coli

High-level production of biologically active human enterokin...

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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： Lixi Niu Jiayue Li Key Laboratory of Chemical Biology and Molecular Engineering of M inistry of Education Shanxi University

Human enterokinase(synonym:enteropeptidase,EC 3.4.21.9)light chain(hEKL)gene wasdesigned and artificially synthesized with built-in codon bias towards Escherichia coli(***)codon *** synthetic hEKL gene was cloned into prokaryotic expressionvector pMAL-s and transferred into the expression strain *** BL21(DE3).The recombinanthEKL protein with a maltose binding protein(MBP)tag was expressed at high levels insoluble form,yielding about 42%of the total cellular *** target protein was thenpurified to the homogeneity better than 95%by affinity *** peptide substrateGST-Melittin with enterokinase recognition site was completely cleaved by the purifiedMBP-hEKL at the molar ratio of 1:5000(enzyme:substrate).Tricine SDS-PAGE analysisshowed that the activity of MBP-hEKL was approximately seven times that of bovine enterokinasecatalytic subunit(EKMaxTM,Invitrogen).From 1 L flask culture,206 mg pure activeMBP-hEKL was obtained and the specific activity was reached to 1.4×104 U/mg.

关键词： Human enterokinase light chain(hEKL) Fusion expression Affinity purification Activity analysis

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Copper complex with ciprofloxacin containing Mannich base hasdual inhibition of Topisomerase

Copper complex with ciprofloxacin containing Mannich base ha...

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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： Yun Fu Yingli Yang Sufeng Zhou Youxun Liu Xingzhi Sun Yanbin Yuan Shaoshan Li changzheng li Xinxiang Medical University

he Mannich base containing ciprofloxacin and kojic acid structural units was prepared andevaluated in antitumor activities. The enhancement in antitumor activity was observedboth from the Mannich base(IC50: 103.3 ± 5.0 μM for HepG2, 87.9 ± 8.0 μM for HCT-116cell) and its copper complex(IC50: 11.5 ± 1.8 μM for HepG2, 44.4 ± 2.5 μM for HCT-116cell) compared to the ciprofloxacin and kojic acid. The mechanistic studies via RT-PCR, cellcycle analysis, mitochondrial membrane potential measurement, inhibition of topoisomeraseand molecular docking indicated that there was different molecular mechanismbetween the Mannich base and its copper complex. The cytotoxicity of Mannich base wasinvolved in apoptosis, cell cycle arrest depolarization of mitochondrial membrane andweaker topoisomerase II inhibition, but the copper complex exerted its cytotoxicity mainlythrough dual topoisomerase inhibition, especially stabilize the intermediate of cleavageDNAtopoisomerase complex.

关键词： Mannich base Copper complex Topisomerase inhibitor

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Comprehensive Profiling of Protein Lysine Acetylation in Escherichia. Coli

Comprehensive Profiling of Protein Lysine Acetylation in Esc...

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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： Zhongyi Cheng PTM Biolabs Inc

Protein lysine acetylation plays a key role in regulating chromatin dynamics, gene expression and metabolic pathways in eukaryotes, and, thus, contributes to diverse cellular processes like transcription, cell cycle regulation, and apoptosis. But the substrates and localization of this modification remain widely unknown due to the limitations of analytical methods. Comprehensive identification of protein acetylation is a major bottleneck due toits dynamic property and pretty low abundance. To solve this problem, we have developed an intergraded approach to identifying lysine acetylation. Combining immunoaffinity enrichment with premium anti-Kac antibodies and high sensitive mass spectrometry analysis(Figure 1), we identified 349 acetylated proteins and addressed 1070 acetylation sites in Escherichiacoli. To our knowledge, the acetylated 21 proteins and acetylated sites were increased to 3 times and 8 times, respectively, compared to that in previous report. These results not only greatly expand the number of acetylated proteins, but also provide a series of important information including localization, networks and characterization of acetylome.

关键词： Protein post-translational modification quantitative proteomics acetylome prokaryote Escherichia.Coli

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Decipher biological significance of the novel epitranscriptomic marker N6-Methyadenosine(m6A) in RNA

Decipher biological significance of the novel epitranscripto...

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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： Yun-Gui Yang CAS Key Laboratory of Genomics and Precision Medicine Beijing Institute of GenomicsChinese Academy of Sciences

The N6-methyladenosine(m6A)modification is one of the most frequently methylated nucleotides in eukaryotic mRNA,accounting for over 80%of all RNA base methylations and present in various *** two independent studies combining m6A-immunoprecipitation with high throughput analysis revealed that the m6A modification tends to mainly occur in intragenic regions including coding sequences(CDS),stop codon flanking regions and 3’-*** frequency was especially high in the 3’-end of CDS and the first quarter of the 3’-UTR,with an average of 1 m6A per 2000 *** m6A modification occurs in highly conserved regions with the consensus sequence:RRACH(R=G or A;H=A,C or U).During the course of brain development a

关键词： in RNA Decipher biological significance of the novel epitranscriptomic marker N6-Methyadenosine m6A

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