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Mitochondria in the pathogenesis of diabetes:a proteomic view

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protein & Cell 2012年第9期3卷 648-660页

作者： Xiulan Chen Shasha Wei Fuquan Yang key laboratory of protein and peptide pharmaceuticals and laboratory of Proteomics Institute of BiophysicsChinese Academy of SciencesBeijing 100101China Graduate University of Chinese Academy of Sciences Beijing 100149China

Diabetes mellitus is a complex metabolic disorder characterized by chronic hyperglycemia due to absolute or relative lack of *** great efforts have been made to investigate the pathogenesis of diabetes,the underlying mechanism behind the development of diabetes and its complications remains *** evidence has linked mitochondrial modification to the pathogenesis of diabetes and its complications and they are also observed in various tissues affected by *** is an attractive tool for the study of diabetes since it allows researchers to compare normal and diabetic samples by identifying and quantifying the differentially expressed proteins in tissues,cells or *** progress has already been made in mitochondrial proteomics to elucidate the role of mitochondria in the pathogenesis of diabetes and its *** studies on the changes of mitochondrial protein specifically post-translational modifications during the diabetic state using proteomic tools,would provide more information to better understand diabetes.

关键词： mitochondria proteomics T1DM T2DM diabetes complication

Oleic Acid and Eicosapentaenoic Acid Reverse Palmitic Acid-induced Insulin Resistance in Human HepG2 Cells via the Reactive Oxygen Species/JUN Pathway

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Genomics, Proteomics & Bioinformatics 2021年第5期19卷 754-771页

作者： Yaping Sun Jifeng Wang Xiaojing Guo Nali Zhu Lili Niu Xiang Ding Zhensheng Xie Xiulan Chen Fuquan Yang key laboratory of protein and peptide pharmaceuticals&laboratory of Proteomics Institute of BiophysicsChinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China

Oleic acid(OA),a monounsaturated fatty acid(MUFA),has previously been shown to reverse saturated fatty acid palmitic acid(PA)-induced hepatic insulin resistance(IR).However,its underlying molecular mechanism is *** addition,previous studies have shown that eicosapentaenoic acid(EPA),aω-3 polyunsaturated fatty acid(PUFA),reverses PA-induced muscle IR,but whether EPA plays the same role in hepatic IR and its possible mechanism involved need to be further ***,we confirmed that EPA reversed PA-induced IR in HepG2 cells and compared the proteomic changes in HepG2 cells after treatment with different free fatty acids(FFAs).A total of 234 proteins were determined to be differentially expressed after PA+OA *** functions were mainly related to responses to stress and endogenous stimuli,lipid metabolic process,and protein *** PA+EPA treatment,the PA-induced expression changes of 1326 proteins could be reversed by EPA,415 of which were mitochondrial proteins,with most of the functional proteins involved in oxidative phosphorylation(OXPHOS)and tricarboxylic acid(TCA)*** studies revealed that the protein encoded by JUN and reactive oxygen species(ROS)play a role in OA-and EPA-reversed PA-induced IR,*** and OA alleviated PA-induced abnormal adenosine triphosphate(ATP)production,ROS generation,and calcium(Ca^(2+))***,H_(2)O_(2)-activated production of ROS increased the protein expression of JUN,further resulting in IR in HepG2 *** together,we demonstrate that ROS/JUN is a common response pathway employed by HepG2 cells toward FFA-regulated IR.

关键词： Free fatty acid Insulin resistance Quantitative proteomics Calcium ATP

Pseudovirus-based neuraminidase inhibition assays reveal potential H5N1 drug-resistant mutations

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protein & Cell 2013年第5期4卷 356-363页

作者： Yi Lu Taijiao Jiang key laboratory of protein and peptide pharmaceuticals National Laboratory of BiomacromoleculesInstitute of BiophysicsChinese Academy of SciencesBeijing 100101China College of Life Sciences University of Chinese Academy of SciencesBeijing 100049China

The use of antiviral drugs such as influenza neuraminidase (NA) inhibitors is a critical strategy to prevent and control flu pandemic, but this strategy faces the challenge of emerging drug-resistant strains. For a highly pathogenic avian influenza (HPAI) H5N1 virus, biosafety restrictions have significantly limited the efforts to monitor its drug responses and mechanisms involved. In this study, a rapid and biosafe assay based on NA pseudovirus was developed to study the resistance of HPAI H5N1 virus to NA inhibitor drugs. The H5N1 NA pseudovirus was comprehensively tested using oseltamivir-sensitive strains and their resistant mutants. Results were consistent with those in previous studies, in which live H5N1 viruses were used. Several oseltamivir-resistant mutations reported in human H1N1 were also identified to cause decreased oseltamivir sensitivity in H5N1 NA by using the H5N1 NA pseudovirus. Thus, H5N1 NA pseudoviruses could be used to monitor HPAI H5N1 drug resistance rapidly and safely.

关键词： infl uenza virus H5N1 neuraminidase inhibi-tor drug resistance pseudovirus

Questions about horse the blood brain barrier receptor 1 spleen ferritin crossing via mouse transferrin

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protein & Cell 2017年第11期8卷 788-790页

作者： Kelong Fan Meng Zhou Xiyun Yan key laboratory of protein and peptide pharmaceuticals CAS-University of Tokyo Joint Laboratory of Structural Virology and Immunology Institute of Biophysics Chinese Academy of Sciences Beijing 100101 China University of Chinese Academy of Sciences Beijing 100049 China

Ferritin, an iron storage protein naturally occurring in the body, has emerged as a promising nanocarrier thanks to its unique architecture, excellent biocompatibility, and ability to self-assemble/disassemble （Fan et al., 2013）. More specifically, the finding that human H-ferritin intrinsically targets tumor cells via binding to its receptor transferrin receptor 1 （TfR1）（Li et al., 2010; Fan et al., 2012; Liang et al., 2014; Zhao et al., 2016） inspired research into using ferritins for tumor target therapy.

关键词： Questions horse the blood brain barrier receptor 1 spleen ferritin crossing via mouse transferrin

TNFR-1 on tumor cells contributes to the sensitivity of fi brosarcoma to chemotherapy

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protein & Cell 2013年第5期4卷 393-401页

作者： Jingjing Deng Xiaopu Zhao Lijie Rong Xiao Li Xiaoman Liu Zhihai Qin key laboratory of protein and peptide pharmaceuticals Chinese Academy of Sciences-University of Tokyo Joint Laboratory of Structural Virology and ImmunologyInstitute of BiophysicsChinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China

Impaired tumor necrosis factor receptor-1(TNFR-1)signaling has been found in some malignant tumors with poor ***,the exact role of TNFR-1 signaling in fi brosarcoma remains ***,we explored the question by comparing the growth of TNFR-1 deficient(Tnfr1-)and TNFR-1 competent(Tnfr1+)fibrosarcoma FB61 cells(FB61-m and FB61-R1)in ***-1 expression on fibrosarcoma cells delayed their growth in vivo but not in ***,reduced FB61-R1 tumor growth was also obtained in T NFR-1 knockout *** mechanism relies mainly on the TNFR-1-mediated down-regulation of vascular endothelial growth factor(VEGF)production by tumor ***,treatment of FB61-m tumors with melphalan resulted in a short delay of tumor growth,followed by a quick r ***,when FB61-R1 tumors were treated with melphalan,tumor growth was similarly delayed at fi rst and then completely *** results reveal evidence for TNFR-1 on tumor cells as a prerequisite in chemotherapy for fi brosarcoma,and provide novel insight into the therapeutic approach against some types of tumors using TNFR-1 angonist.

关键词： TNFR-1 fi brosarcoma chemotherapy

A new method for improving the accuracy of miRNA detection with NaYF4：Yb,Er upconversion nanoparticles

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Science China Chemistry 2017年第1期60卷 157-162页

作者： Lanlan Mao Zhuoxuan Lu Nongyue He Liming Zhang Yan Deng Deming Duan Economical Forest Cultivation and Utilization of 2011 Collaborative Innovation Center in Hunan Province Hunan Key Laboratory of Green Packaging and Application of Biological Nanotechnology Hunan University of Technology Zhuzhou 412007 China key laboratory of protein and peptide pharmaceuticals Institute of Biophysics Chinese Academy of Sciences Beijing 100101 China State key laboratory of Bioelectronics School of Biological Science and Medical Engineering Southeast University Nanjing 210096 China

MicroRNAs(miRNAs)are small noncoding RNAs,which play a central role in gene expression regulation and have been considered as excellent biomarker candidates for clinical diagnosis and *** far,many miRNAs detection methods require polymerase chain reaction(PCR)amplification following reverse transcription of *** processes are complicated and *** this work,we have developed a simpler method for miRNA detection based on base stacking hybridization happening on the surface of NaYF_4:Yb,Er upconversion *** this method,the fluorescence of NaYF_4:Yb,Er upconversion nanoparticles were functionalized as a reference standard,which can improve the accuracy of miRNA *** the basis of these findings,we suggest this novel approach for miRNA detection could be applied as an accurate and specific technique for miRNAs detection.

关键词： microRNAs detection NaYF4:Yb Er reference standard base stacking hybridization

Affinity maturation of anti-TNF-alpha scFv with somatic hypermutation in non-B cells

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protein & Cell 2012年第6期3卷 460-469页

作者： Shaopeng Chen Junkang Qiu Chuan Chen Chunchun Liu Yuheng Liu Lili An Junying Jia Jie Tang Lijun Wu Haiying Hang key laboratory for protein and peptide pharmaceuticals National Laboratory of BiomacromoleculesInstitute of BiophysicsChinese Academy of SciencesBeijing 100101China key laboratory of Ion Beam Bioengineering Institute of Technical Biology and Agriculture EngineeringChinese Academy of SciencesHefei 230031China Core Facility Center Institute of BiophysicsChinese Academy of SciencesBeijing 100101China􀀍

Activation-induced cytidine deaminase(AID)is required for the generation of antibody diversity through initiat-ing both somatic hypermutation(SHM)and class switch recombination.A few research groups have success-fully used the feature of AID for generating mutant li-braries in directed evolution of target proteins in B cells in vitro.B cells,cultured in suspension,are not con-venient for transfection and *** this study,we established an AID-based mutant accumulation and sorting system in adherent human *** AID gene was first transfected into the human non-small cell lung carcinoma H1299 cells,and a stable cell clone(H1299-AID)was ***,anti-hTNF-αscFv(ATscFv)was transfected into H1299-AID cells and ATscFv was displayed on the surface of H1299-AID *** 4-round amplification/flow cytometric sorting for cells with the highest affinities to hTNF-alpha,two ATscFv mutant gene clones were *** with the wild type ATscFv,the two mutants were much more efficient in neutralizing cytotoxicity of *** results indicate that directed evolution by somatic hypermutation can be carried out in adherent non-B cells,which makes directed evolution in mammalian cells easier and more efficient.

关键词： antibody activation-induced cytidine deaminase(AID) somatic hypermutation affinity maturation TNF-alpha

Nanozymes： an emerging field bridging nanotechnology and biology

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Science China(Life Sciences) 2016年第4期59卷 400-402页

作者： Lizeng Gao Xiyun Yan key laboratory of protein and peptide pharmaceuticals CAS-University of Tokyo Joint Laboratory of Structural Virology and Immunology Institute of Biophysics Chinese Academy of Sciences Beijing 100101 China Jiangsu key laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases School of Medicine Yangzhou University Yangzhou 225001 China

Enzymes are biological catalysts that can convert substrates into products in biochemical *** 1926,the first enzyme,urease,was determined to be a protein by James *** who won the Nobel Prize in *** then,enzymes have been considered to be proteins,which allows them to achieve their high catalytic activity with high specific activity under mild ***,in general,the enzyme activity of proteins is lost after exposure to extremes of p H and high temperature,and proteins are also susceptible to digestion by proteases in the environment,which dramatically hinders their practical applications in

关键词： nanotechnology bridging digestion biochemical emerging exposure convert susceptible dramatically enzymes

10E8-like neutralizing antibodies against HIV-1 induced using a precisely designed conformational peptide as a vaccine prime

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Science China(Life Sciences) 2014年第1期57卷 117-127页

作者： YU Yang TONG Pei LI Yu LU ZhiFeng CHEN YingHua laboratory of Immunology School of Life Sciences Tsinghua University Beijing Key Laboratory for Protein Therapeutics MOE key laboratory for protein Sciences

Recent studies have demonstrated that the membrane-proximal external region （MPER） of human immunodeficiency virus 1 （HIV-1） glycoprotein 41 contains a series of epitopes for human monoclonal antibodies, including 2F5, Zl3el, 4El0, and 10E8, which were isolated from HIV-l-infected individuals and show broad neutralizing activities. This suggests that MPER is a good target for the development of effective HIV-1 vaccines. However, many studies have shown that it is difficult to induce antibodies with similar broad neutralizing activities using MPER-based peptide antigens. Here, we report that 10E8-1ike neu- tralizing antibodies with effective anti-HIV-1 activity were readily induced using a precisely designed conformational immu- nogenic peptide containing the 10E8-specific epitope. This immunogenic peptide （designated T10HE） contains a 15-mer MPER-derived 10E8-specific epitope fused to T-helper-cell epitopes from tetanus toxin （tt）, which showed a significantly sta- bilized a-helix structure after a series of modifications, including substitution with an （S）-c^-（2＇-pentenyl） alanine containing an olefin-bearing tether and ruthenium-catalyzed olefin metathesis, compared with the unmodified T10E peptide. The stabilized （x-helix structure of T10HE did not affect its capacity to bind the 10E8 antibody, as evaluated with an enzyme linked immuno- sorbent assay （ELISA） and surface plasmon resonance binding assay （SPR assay）. The efficacies of the T10HE and T10E epitope vaccines were evaluated after a standard vaccination procedure in which the experimental mice were primed with ei- ther the T10HE or T10E immunogen and boosted with HIV-1 JRFL pseudoviruses. Higher titers of 10E8-1ike antibodies were induced by T10HE than that by T10E. More importantly, the antibodies induced by T10HE showed enhanced antiviral potency against HIV-1 strains with both X4 and R5 tropism and a greater degree of broad neutralizing activity than the antibodies in- duced by T10E. These results indic

关键词： MPER peptide immunogen vaccine prime neutralizing antibody