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Comprehensive analysis of the N and C terminus of endogenous serum peptides reveals a highly conserved cleavage site pattern derived from proteolytic enzymes

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Protein & Cell 2012年第9期3卷 669-674页

作者： fangjun wang Jun Zhu Lianghai Hu Hongqiang Qin Mingliang Ye Hanfa Zou CAS Key Lab of Separation Sciences for Analytical Chemistry National Chromatographic Research and Analysis CenterDalian Institute of Chemical PhysicsChinese Academy of SciencesDalian 116023China

The human serum proteome is closely associated with the state of the *** peptides derived from proteolytic enzymes cleaving on serum proteins are widely studied due to their potential application in disease-specific marker ***,the reproducibility of peptidome analysis of endogenous peptides is significantly influenced by the proteolytic enzymes within body fluids,thereby limiting the clinical use of the endogenous *** comprehensively investigated the N and C terminus of endogenous peptides using *** cleavage site patterns of the N and C terminus and adjacent sites from all the identified endogenous peptides were highly conserved under different sample preparation conditions,including long-term incubation at 37℃ and pretreatment with repeated freeze-thaw ***,a distinguishable cleavage site pattern was obtained when a different disease serum was *** conserved cleavage site pattern derived from proteolytic enzymes holds potential in highly specific disease diagnosis.

关键词： human serum endogenous peptide N and C termini disease diagnosis

Ion-pair Reversed-phase×Low-pH Reversed-phase Two-dimensional Liquid Chromatography for In-depth Proteomic Profiling

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Chemical Research in Chinese Universities 2023年第2期39卷 260-265页

作者： NIU Wenxue LIU Zheyi LIU Jing LAI Can ZHANG Tingting ZHAO Heng wang Guosheng wang fangjun College of Chemical Engineering Shenyang University of Chemical TechnologyShenyang110142P.R.China CAS Key Laboratory of Separation Sciences for Analytical Chemistry Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian116023P.R.China College of Pharmacy Dalian Medical UniversityDalian116044P.R.China University of Chinese Academy of Sciences Beijing100049P.R.China

High-resolution liquid chromatography separation is essential to in-depth proteomic profiling of complex biological ***,we established an ion-pair reversed-phase×reversed-phase two-dimensional liquid chromatography(IPRP×RP 2DLC)strategy for comprehensive proteomic *** RPLC separation dimensions were performed at low pH,with trifluoroacetic acid(TFA)and formic acid(FA)as mobile phase addictive,*** the good separation resolution offered by ion-pairing effect of TFA,the fractionation efficiency was greatly improved with 74.0%peptides identified in just one *** with conventional high pH RP fractionation,the overall separation rate of IPRP was about 1.6 times that of high-pH RP,which increased the number of identified peptides by 21%.Further,2169 proteins and 8540 peptides were confidently identified from crude serum sample by our IPRP×RP 2DLC strategy,exhibiting great potential in clinical proteomics in the future.

关键词： Two-dimensional liquid chromatography(2DLC) Ion-pair reversed-phase liquid chromatography(IPRP-LC) IPRP×low-pH RP 2DLC Proteomics Mass spectrometry

Characterization and manipulation of the photosystem Ⅱ-semiconductor interfacial molecular interactions in solar-to-chemical energy conversion

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Journal of Energy Chemistry 2022年第7期31卷 437-443,I0012页

作者： Min He wangyin wang Zheyi Liu Wenxiang Zhang Jinan Li Wenming Tian Ye Zhou Yan Jin fangjun wang Can Li CAS Key Laboratory of Separation Sciences for Analytical Chemistry Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian 116023LiaoningChina University of Chinese Academy of Sciences Beijing 100049China State Key Laboratory of Catalysis Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian National Laboratory for Clean EnergyDalian 116023LiaoningChina State Key Laboratory of Molecular Reaction Dynamics Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian 116023LiaoningChina

Semi-artificial photosynthesis interfacing catalytic protein machinery with synthetic photocatalysts exhibits great potential in solar-to-chemical energy conversion. However, characterizing and manipulating the molecular integration structure at the biotic-abiotic interface remain a challenging task. Herein,the biointerface molecular integration details of photosystem II(PSII)-semiconductor hybrids, including the PSII orientation, interfacial microdomains, and overall structure modulation, are systematically interrogated by lysine reactivity profiling mass spectrometry. We demonstrate the semiconductor surface biocompatibility is essential to the PSII self-assembly with uniform orientation and electroactive *** directional localization of PSII onto more hydrophilic Ru/Sr Ti O_(3):Rh surface exhibits less disturbance on PSII structure and electron transfer chain, beneficial to the high water splitting ***, rational modification of hydrophobic Ru_(2)S_(3)/Cd S surface with biocompatible protamine can improve the hybrid O_(2)-evolving activity 83.3%. Our results provide the mechanistic understanding to the structure–activity relationship of PSII-semiconductor hybrids and contribute to their rational design in the future.

关键词： PSII-semiconductor hybrids Biointerface molecular interaction Solar-to-chemical energy conversion Mass spectrometry Lysine reactivity profiling

Catalyst-free Photochemical Bromination of Unprotected Aromatic Amino Acid Derivatives by Using a Rotating Ultraviolet Photoreactor

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Chemical Research in Chinese Universities 2022年第2期38卷 505-509页

作者： ZHANG Tingting LUO Pan LAI Can LIU Zheyi JIN Yan wang fangjun CAS Key Laboratory of Separation Sciences for Analytical Chemistry Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian 116023P.R.China University of Chinese Academy of Sciences Beijing 100049P.R.China

Aromatic amino acid halides are valuable amino acid derivatives and fundamental building blocks that are used to various coupling reaction^1-3!,such as Heck,Suzuki and Ullmann reaction[4-7],and also important precurso... 详细信息

关键词： Catalyst-free Photochemistry Bromination 254 nm UV Unprotected aromatic amino acid derivative

Elucidating the various multi-phosphorylation statuses of protein functional regions by 193-nm ultraviolet photodissociation

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Chinese Chemical Letters 2018年第5期29卷 694-698页

作者： Zheyi Liu You Jiang Chunlei Xiao Xingchuang Xiong Tao wang Jinan Li fangjun wang Xiang Fang Xueming Yang CAS Key Laboratory of Separation Sciences for Analytical Chemistry Dalian Institute of Chemical Physics Chinese Academy of Sciences Dalian 116023 China National Institute of Metrology Beijing 100012 China State Key Laboratory of Molecular Reaction Dynamics Dalian Institute of Chemical Physics Chinese Academy of Sciences Dalian 116023 China

Ultraviolet photodissociation is a high-energy fast excitation method in mass spectrometry and has beensuccessfully applied for the elucidation of sequences and structures of biomolecules. However, its abilityto distinguish the phosphorylation sites isomers of multi-phosphopeptides has been not systematicallyinvestigated until now. A 193-nm ultraviolet laser dissociation mass spectrometry system wasestablished in this study and applied to elucidate the complex multi-phosphorylation statuses mimickingthe functional regions of Sicl, Gli3 and Tau. The numbers of matched fragment ions and phosphorylationsite-determining ions were improved on average 123% and 104%, respectively, by utilizing the ultravioletphotodissociation strategy, comparing to the typically utilized collision induced dissociation ***. 94% phosphorylation sites within various statuses were unambiguously elucidated.

关键词： Ultraviolet photodissociation Mass spectrometry Multi-phosphorylation Phosphorylated tyrosine Phosphorylation site-determining ion

In-situ generation and global property profiling of metal nanoclusters by ultraviolet laser dissociation-mass spectrometry

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Science China Chemistry 2022年第6期65卷 1196-1203页

作者： Zheyi Liu Zhaoxian Qin Chaonan Cui Zhixun Luo Bing Yang You Jiang Can Lai Zhipeng wang Xiaolei wang Xiang Fang Gao Li fangjun wang Chunlei Xiao Xueming Yang State Key Laboratory of Molecular Reaction Dynamics Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian116023China CAS Key Laboratory of Separation Sciences for Analytical Chemistry Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian116023China State Key Laboratory of Catalysis Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian116023China State Key Laboratory for Structural Chemistry of Unstable and Stable Species Institute of ChemistryChinese Academy of SciencesBeijing100190China Dalian National Laboratory for Clean Energy Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian116023China National Institute of Metrology Beijing100013China University of Chinese Academy of Sciences Beijing100049China

Metal nanoclusters are promising nanomaterials with unique properties, but only a few ones with specific numbers of metal atoms can be obtained and studied up to now. In this study, we establish a new paradigm of in-situ generation and global study of metal nanoclusters with different sizes, constitutions, and charge states, including both accurate constitution characterization and global activity profiling. The complex mixtures of metal nanoclusters are produced by employing single-pulsed 193-nm laser dissociation of monolayer-protected cluster(MPC) precursors within a high-resolution mass spectrometry(HRMS). More than400 types of bare gold nanoclusters including novel multiply charged(2+ and 3+), S-/P-doped, and silver alloy ones can be efficiently generated and accurately characterized. A distinct size(1 to 142 atoms)-and charge(1+ to 3+)-hierarchy reactivity is clearly observed for the first time. This global cluster study might greatly promote the developments and applications of novel metal nanoclusters.

关键词： metal nanocluster ultraviolet laser dissociation mass spectrometry global property profiling size-and charge-hierarchy

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Highly efficient artificial blood coagulation shortcut confined on Ca-zeolite surface

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Nano Research 2021年第9期14卷 3309-3318页

作者： Lisha Yu Bin Yu Hao Chen Xiaoqiang Shang Min He Mengchi Lin Dan Li Wenzhao Zhang Zhengzhong Kang Jiachen Li fangjun wang Liping Xiao Qi wang Jie Fan Department of Chemistry Zhejiang UniversityHangzhou 310027China CAS Key Laboratory of Separation Sciences for Analytical Chemistry Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian 116023China

It is challenging to develop an in w'fro catalytic system to conduct natural surface-confined enzymatic reactions in a stable,efficient,and spatially defined ***,we report that an artificial catalyst,which composes of trypsin and a calcium ion exchanged zeolite Y(trypsin/CaY),is capable of conducting surface-confined thrombin generation,and then constructs an artificial shortcut for classic,natural and complex blood coagulation *** Ca2+within the microporous cages play a key role in trypsin/CaY hybrid through tuning the bio-inorganic interaction and spatial orientation of the protease,which allows trypsin/CaY to display greatly enhanced catalytic performance in coagulation *** in vivo efficiency of the artificial coagulation shortcut is further confirmed in massive bleeding and hemophilia animal *** hemostasis is achieved by trypsin/CaY hybrid in a hemophilia A mice tail bleeding model,where natural clotting system fails in response to bleeding event due to factor VIII *** a rabbit lethal femoral artery injury model,the blood loss of the artificial catalyst is decreased by 4-7 fold when compared to state-of-art clay-or zeolite-based topical agents.

关键词： surface-confined Ca-zeolite blood coagulation trypsin

Elucidating the Molecular Mechanism of Dynamic Photodamage of Photosystem II Membrane Protein Complex by Integrated Proteomics Strategy

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CCS Chemistry 2022年第1期4卷 182-193页

作者： Ye Zhou Zheyi Liu Mingdong Yao Jun Chen Yanan Xiao Guangye Han Jian-Ren Shen fangjun wang CAS Key Laboratory of Separation Sciences for Analytical Chemistry Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian 116023 State Key Laboratory of Catalysis Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian 116023 Key Laboratory of Systems Bioengineering Ministry of EducationTianjin UniversityTianjin 300072 Science and Technology on Surface Physics and Chemistry Laboratory Jiangyou 621908 University of Chinese Academy of Sciences Beijing 100049 Photosynthesis Research Center Key Laboratory of PhotobiologyInstitute of BotanyChinese Academy of SciencesBeijing 100093 Research Institute for Interdisciplinary Science Graduate School of Natural Science and TechnologyOkayama UniversityOkayama 700-8530

Photosystem II(PSII),as a multiple-subunit chloroplast membrane-associated pigment-protein complex on the thylakoid membrane,is a primary target of light-induced ***,the overall molecular details of the conformation and composition dynamics of PSII photodamage are still *** this study,we investigated systematically the dynamic conformation,degradation,and oxidation processes of PSII photodamage by integrating chemical cross-linking and top-down proteomics strategies.

关键词： photosystem II dynamic photodamage chemical cross-linking top-down proteomics mass spectrometry

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中国上市公司研发支出的非线性调整研究

系统工程理论与实践 2018年第12期38卷 3050-3058页

作者：汪方军孙俊勤王璇子西安交通大学管理学院西安710049 西安交通大学管理国家级实验教学示范中心西安710049

本文以2011—2016年中国上市公司为研究对象,建立了企业研发支出与主营业务收入之间关系的计量经济模型.研究发现研发支出调整与主营业务收入变动之间存在非线性关系,证实了研发支出存在粘性特征:当主营业务收入上升1%时,研发支出增加0.... 详细信息

本文以2011—2016年中国上市公司为研究对象,建立了企业研发支出与主营业务收入之间关系的计量经济模型.研究发现研发支出调整与主营业务收入变动之间存在非线性关系,证实了研发支出存在粘性特征:当主营业务收入上升1%时,研发支出增加0.634%;而主营业务收入下降1%时,研发支出减少0.401%.进一步研究发现,当主营业务收入连续两年下降时,研发支出粘性会降低;研发支出粘性也会随着宏观经济增长幅度、劳动密集程度增加而增大;与民营企业相比较,国有企业的研发支出粘性更大.研发支出粘性的研究有助于揭示企业管理层研发支出调整“黑箱”.

关键词：研发支出主营业务收入非线性调整粘性