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Enzymatic self-assembly/disassembly turns “ON”/“OFF” the mimetic hydrolytic activity of histidine nanofibers

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Science China Chemistry 2024年第2期67卷 517-522页

作者： Xianbao Sun manli wang Xiaoyang Liu Wenjun Zhan Guangqiong Xu Qiaochu Jiang Hai-Dong Xu Fuqiang wang Gaolin Liang State Key Laboratory of Digital Medical Engineering School of Biological Science and Medical EngineeringSoutheast UniversityNanjing210096China Analysis Center Nanjing Medical UniversityNanjing210029China

Mimetic enzymes are devised as alternates or supplements of natural enzymes in broad fields but regulating their activities in a switchable manner remains challenging.Herein,we proposed an enzymatic self-assembly/disassembly strategy to address this issue.A peptide molecule Nap FFEYIH(YH) was rationally designed which,after self-assembling into nanofibers,lined up the histidine moieties to form active hydrolysis centers for mimicking hydrolase activity.Enzymatic dephosphorylation of Nap FFEYp IH(Yp H) by alkaline phosphatase to yield YH also turned “ON” the hydrolase activity.In turn,phosphorylation of YH by phosphokinase epidermal growth factor receptor to yield Yp H disassembled the nanofibers and thus turned the activity “OFF”.As such,the “ON”/“OFF” of the mimetic hydrolase activities could be regulated under physiological conditions through ALP/EGFR-mediated self-assembly/disassembly of histidine nanofibers.This work provides a feasible strategy for the on-demand fabrication of artificial enzymes with controllable and superior activities.

关键词： disassembly hydrolytic activity mimetic enzyme phosphorylation self-assembly

Structural Isomers:Small Change with Big Difference in Anion Storage

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Nano-Micro Letters 2024年第1期16卷 222-234页

作者： Huichao Dai Yuan Chen Yueyue Cao manli Fu Linnan Guan Guoqun Zhang Lei Gong Mi Tang Kun Fan Chengliang wang School of Integrated Circuits Wuhan National Laboratory for Optoelectronics(WNLO)Huazhong University of Science and TechnologyWuhan 430074People’s Republic of China Wenzhou Advanced Manufacturing Institute Huazhong University of Science and TechnologyWenzhou 325035People’s Republic of China Hubei Key Laboratory of Polymer Materials School of Materials Science and EngineeringHubei UniversityWuhan 430062People’s Republic of China School of Chemistry and Environmental Engineering Wuhan Institute of TechnologyWuhan 430073People’s Republic of China

Organic electrode materials are promising for batteries.However,the reported organic electrodes are often facing the challenges of low specific capacity,low voltage,poor rate capability and vague charge storage mechanisms,etc.Isomers are good platform to investigate the charge storage mechanisms and enhance the performance of batteries,which,however,have not been focused in batteries.Herein,two isomers are reported for batteries.As a result,the isomer tetrathiafulvalene(TTF)could store two monovalent anions reversibly,deriving an average discharge voltage of 1.05 V and a specific capacity of 220 mAh g−1 at a current density of 2 C.On the other hand,the other isomer tetrathianaphthalene could only reversibly store one monovalent anion and upon further oxidation,it would undergo an irreversible solid-state molecular rearrangement to TTF.The molecular rearrangement was confirmed by electrochemical performances,X-ray diffraction patterns,nuclear magnetic resonance spectra,and 1H detected heteronuclear multiple bond correlation spectra.These results suggested the small structural change could lead to a big difference in anion storage,and we hope this work will stimulate more attention to the structural design for boosting the performance of organic batteries.

关键词： Zinc-organic batteries Isomers Solid-state molecular rearrangement Anion storage P-type organic electrode materials

Factors Influencing Alzheimer's Disease Risk: Whether and How They are Related to the APOE Genotype

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Neuroscience Bulletin 2022年第7期38卷 809-819页

作者： Rong Zhang Xiaojiao Xu Hang Yu Xiaolan Xu manli wang Weidong Le Institute of Neurology Sichuan Academy of Medical SciencesSichuan Provincial People's HospitalUniversity of Electronic Science and Technology of ChinaChengdu 611731China

Alzheimer's disease(AD)is the most prevalent neurodegenerative disease featuring progressive cognitive impairment.Although the etiology of late-onset AD remains unclear,the close association of AD with apolipoprotein E(APOE),a gene that mainly regulates lipid metabolism,has been firmly established and may shed light on the exploration of AD pathogenesis and therapy.However,various confounding factors interfere with the APOE-related AD risk,raising questions about our comprehension of the clinical findings concerning APOE.In this review,we summarize the most debated factors interactingwith theAPOEgenotype and ADpathogenesis,depict the extent to which these factors relate to APOEdependent AD risk,and discuss the possible underlying mechanisms.

关键词： Apolipoprotein E Alzheimer's disease Ethnicity Diet Geographic factor Aging Gender

Genomics and proteomics of Apis mellifera filamentous virus isolated from honeybees in China

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Virologica Sinica 2022年第4期37卷 483-490页

作者： Dahe Yang Jun wang Xi wang Fei Deng Qingyun Diao manli wang Zhihong Hu Chunsheng Hou Institute of Apicultural Research Chinese Academy of Agricultural SciencesBeijing100093China State Key Laboratory of Virology and National Virus Resource Center Wuhan Institute of VirologyCenter for Biosafety Mega-ScienceChinese Academy of SciencesWuhan430071China Institute of Bast Fiber Crops Chinese Academy of Agricultural SciencesChangsha410205China

Apis mellifera filamentous virus(Am FV)is a large DNA virus that is endemic in honeybee colonies.The genome sequence of the Am FV Swiss isolate(Am FV CH–C05)has been reported,but so far very few molecular studies have been conducted on this virus.In this study,we isolated and purified Am FV(Am FV CN)from Chinese honeybee(Apis mellifera)colonies and elucidated its genomics and proteomics.Electron microscopy showed ovoid purified virions with dimensions of 300–500×210–285 nm,wrapping a 3165×40 nm filamentous nucleocapsid in three figure-eight loops.Unlike Am FV CH–C05,which was reported to have a circular genome,our data suggest that Am FV CN has a linear genome of approximately 493 kb.A total of 197 ORFs were identified,among which36 putative genes including 18 baculoviral homologs were annotated.The overall nucleotide similarity between the CN and CH–C05 isolates was 96.9%.Several ORFs were newly annotated in Am FV CN,including homologs of per os infectivity factor 4(PIF4)and a putative integrase.Phylogenomic analysis placed Am FVs on a separate branch within the newly proposed virus class Naldaviricetes.Proteomic analysis revealed 47 Am FV virionassociated proteins,of which 14 had over 50%sequence coverage,suggesting that they are likely to be main structural proteins.In addition,all six of the annotated PIFs(PIF-0–5)were identified by proteomics,suggesting that they may function as entry factors in Am FV infection.This study provides fundamental information regarding the molecular biology of Am FV.

关键词： Apis mellifera filamentous Virus(AmFV) per os infectivity factor 4(PIF4) Genome sequence Proteomics Structural proteins Naldaviricetes

Construction and characterization of a synthesized herpes simplex virus H129-Syn-G2

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Virologica Sinica 2023年第3期38卷 373-379页

作者： Han Xiao Hengrui Hu Yijia Guo Jiang Li Le Wen Wen-Bo Zeng manli wang Min-Hua Luo Zhihong Hu State Key Laboratory of Virology Center for Biosafety Mega-ScienceWuhan Institute of VirologyChinese Academy of SciencesWuhan430071China University of Chinese Academy of Sciences Beijing100049China

Herpes simplex virus type 1(HSV-1)causes lifelong infections worldwide,and currently there is no efficient cure or vaccine.HSV-1-derived tools,such as neuronal circuit tracers and oncolytic viruses,have been used exten-sively;however,further genetic engineering of HSV-1 is hindered by its complex genome structure.In the present study,we designed and constructed a synthetic platform for HSV-1 based on H129-G4.The complete genome was constructed from 10 fragments through 3 rounds of synthesis using transformation-associated recombination(TAR)in yeast,and was named H129-Syn-G2.The H129-Syn-G2 genome contained two copies of the gfp gene and was transfected into cells to rescue the virus.According to growth curve assay and electron microscopy results,the synthetic viruses exhibited more optimized growth properties and similar morphogenesis compared to the parental virus.This synthetic platform will facilitate further manipulation of the HSV-1 genome for the devel-opment of neuronal circuit tracers,oncolytic viruses,and vaccines.

关键词： Herpes simplex virus type 1(HSV-1) Neuronal circuit tracers H129-Syn-G2 H129-G4 Synthetic biology

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画史互鉴——长安画派作品中的人民形象

美术 2024年第3期 141-143页

作者：王欣王曼利西北大学文学院西安中国画院咸阳师范学院

长安画派以“人民”为核心表现对象,对“劳动着的人民”的捕捉传递了积极乐观、坚韧顽强的精神风貌;通过描绘少数民族群体的安定团结反映“民族一家亲”的民族政策;在人民当家做主的语境下彰显女性主体意识的觉醒。长安画派对人民形象... 详细信息

长安画派以“人民”为核心表现对象,对“劳动着的人民”的捕捉传递了积极乐观、坚韧顽强的精神风貌;通过描绘少数民族群体的安定团结反映“民族一家亲”的民族政策;在人民当家做主的语境下彰显女性主体意识的觉醒。长安画派对人民形象的书写,不仅塑造了中华人民共和国建设时期的民族形象,也奠定了长安画派在中国美术史上的重要地位,展现出画史互鉴的深刻意义。

关键词：长安画派劳动人民少数民族劳动女性

Hybridoma-derived neutralizing monoclonal antibodies against Beta and Delta variants of SARS-CoV-2 in vivo

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Virologica Sinica 2023年第2期38卷 257-267页

作者： Qianran wang Lu Peng Yanqiu Nie Yanni Shu Huajun Zhang Zidan Song Yufeng Li Hengrui Hu Liushuai Li Xi wang Jia Liu Jiang Li Zhengli Shi Fei Deng Yu Guo Yiwu Zhou Bing Yan Zhihong Hu manli wang State Key Laboratory of Virology Wuhan Institute of VirologyCenter for Biosafety Mega-ScienceChinese Academy of SciencesWuhan430071China State Key Laboratory of Medicinal Chemical Biology College of Life SciencesNankai UniversityTianjin300071China Department of Forensic Medicine Tongji Medical College of Huazhong University of Science and TechnologyWuhan430010China Hubei Jiangxia Laboratory Wuhan430200China

Neutralizing monoclonal antibodies(mAb)are a major therapeutic strategy for the treatment of severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)infection.The continuous emergence of new SARS-CoV-2 variants worldwide has increased the urgency for the development of new mAbs.In this study,we immunized mice with the receptor-binding domain(RBD)of the SARS-CoV-2 prototypic strain(WIV04)and screened 35 RBDspecific mAbs using hybridoma technology.Results of the plaque reduction neutralization test showed that 25 of the mAbs neutralized authentic WIV04 strain infection.The 25 mAbs were divided into three categories based on the competitive enzyme-linked immunosorbent assay results.A representative mAb was selected from each category(RD4,RD10,and RD14)to determine the binding kinetics and median inhibitory concentration(IC_(50))of WIV04 and two variants of concern(VOC):B.1.351(Beta)and B.1.617.2(Delta).RD4 neutralized the B.1.617.2 variant with an IC50 of 2.67 ng/mL;however,it completely lost neutralizing activity against the B.1.351 variant.RD10 neutralized both variants with an IC50 exceeding 100 ng/mL;whereas RD14 neutralized two variants with a higher IC50(>1 mg/mL).Animal experiments were performed to evaluate the protective effects of RD4 and RD10 against various VOC infections.RD4 could protect Adv-hACE2 transduced mice from B.1.617.2 infection at an antibody concentration of 25 mg/kg,while RD10 could protect mice from B.1.351 infection at an antibody concentration of 75 mg/kg.These results highlight the potential for future modifications of the mAbs for practical use.

关键词： COVID-19 SARS-CoV-2 B.1.351 B.1.617.2 Monoclonal antibody

An in vitro recombination-based reverse genetic system for rapid mutagenesis of structural genes of the Japanese encephalitis virus

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Virologica Sinica 2015年第5期30卷 354-362页

作者： Ruikun Du manli wang Zhihong Hu Hualin wang Fei Deng State Key Laboratory of Virology Wuhan Institute of VirologyChinese Academy of Sciences

Japanese encephalitis virus(JEV) is one of the most common pathogens of severe viral encephalitis, which is a severe threat to human health. Despite instability of the JEV genome in bacteria, many strategies have been developed to establish molecular clone systems of JEV, providing convenient tools for studying the virus life cycle and virus–host interactions. In this study, we adapted an In-Fusion enzyme-based in vitro recombination method to construct a reverse genetic system of JEV, thereby providing a rapid approach to introduce mutations into the structural genes. A truncated genome without the structural genes was constructed as the backbone, and the complementary segment containing the structural genes was recombined in vitro, which was then transfected directly into virus-permissive cells. The progeny of the infectious virus was successfully detected in the supernatant of the transfected cells, and showed an identical phenotype to its parental virus. To provide a proof-of-principle, the 12 conserved cysteine residues in the envelope(E) protein of JEV were respectively mutated using this approach, and all mutations resulted in a complete failure to generate infectious virus. However, a leucine-tophenylanine mutation at amino acid 107 of the E protein did not interfere with the production of the infectious virus. These results suggested that all 12 cysteines in the E protein are essential for the JEV life cycle. In summary, a novel reverse genetic system of JEV was established for rapidly introducing mutations into structural genes, which will serve as a useful tool for functional studies.

关键词： Japanese encephalitis virus(JEV) reverse genetic s

Improving Baculovirus Transduction of Mammalian Cells by Incorporation of Thogotovirus Glycoproteins

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Virologica Sinica 2019年第4期34卷 454-466页

作者： Liangbo Hu Yimeng Li Fei Deng Zhihong Hu Hualin wang manli wang State Key Laboratory of Virology Wuhan Institute of VirologyChinese Academy of SciencesWuhan 430071China University of the Chinese Academy of Sciences Beijing 100049China

Baculovirus can transduce a wide range of mammalian cells and is considered a promising gene therapy vector. However,the low transduction efficiency of baculovirus into many mammalian cells limits its practical application. Co-expressing heterologous viral glycoproteins(GPs), such as vesicular stomatitis virus G protein(VSV G), with baculovirus native envelope protein GP64 is one of the feasible strategies for improving virus transduction. Tick-borne thogotoviruses infect mammals and their GPs share sequence/structure homology and common evolutionary origins with baculovirus GP64.Herein, we tested whether thogotovirus GPs could facilitate the entry of the prototype baculovirus Autographa californica multiple multiple nucleopolyhedrovirus(AcMNPV) into mammalian cells. The gp genes of two thogotoviruses, Thogoto virus and Dhori virus, were inserted into the AcMNPV genome. Both GPs were properly expressed and incorporated into the envelope of the recombinant AcMNPVs. The transduction rates of recombinant AcMNPVs expressing the two thogotovirus GPs increased for approximately 4–12 fold compared to the wild type AcMNPV in six of the 12 tested mammalian cell lines. It seemed that thogotovirus GPs provide the recombinant AcMNPVs with different cell tropisms and showed better performance in several mammalian cells compared to VSV G incorporated AcMNPV. Further studies showed that the improved transduction was a result of augmented virus-endosome fusion and endosome escaping, rather than increased cell binding or internalization. We found the AcMNPV envelope protein GP64-mediated fusion was enhanced by the thogotovirus GPs at relatively higher p H conditions. Therefore, the thogotovirus GPs represent novel candidates to improve baculovirus-based gene delivery vectors.

关键词： Autographa californica multiple nucleopolyhedrovirus(AcMNPV) Baculovirus Thogotovirus Glycoprotein Transduction Mammalian cells