综述了G蛋白偶联受体(G protein coupled receptor,GPCR)二聚化检测的生物化学、生物物理、结构生物学方法,系统阐述了相关原理与关键要点,并对比分析了优势与局限性。未来研究应聚焦于开发更加灵敏且非侵入性的GPCR二聚化检测方法,推...综述了G蛋白偶联受体(G protein coupled receptor,GPCR)二聚化检测的生物化学、生物物理、结构生物学方法,系统阐述了相关原理与关键要点,并对比分析了优势与局限性。未来研究应聚焦于开发更加灵敏且非侵入性的GPCR二聚化检测方法,推动多种技术手段的协同联用,从而深入探索GPCR二聚体在疾病发生和发展中的作用,为药物设计提供新思路,并进一步发掘其在作物病虫害防治与畜牧业的应用潜力。展开更多
Novel insights into complex biological processes very often critically depend on the establishment of new potent read-out tools and improved protocols.A lot has been learned over the past four decades on physiological...Novel insights into complex biological processes very often critically depend on the establishment of new potent read-out tools and improved protocols.A lot has been learned over the past four decades on physiological functions and,importantly,disease-related roles of the prion protein(PrP),a relatively broadly expressed membrane-anchored glycoprotein with high levels in several cell types of the nervous and immune system and with well-established key roles in different progressive and fatal neurodegenerative protein misfolding diseases(proteopathies).展开更多
The study of target proteins is crucial for understanding molecular interactions and developing analytical platforms,therapeutic agents and functional tools.Herein,we present a novel nanoplatform activated by near-inf...The study of target proteins is crucial for understanding molecular interactions and developing analytical platforms,therapeutic agents and functional tools.Herein,we present a novel nanoplatform activated by near-infrared(NIR) light for triple-modal proteins study,which enabling target protein labeling,enrichment and visualization.Azido-naphthalimide-coated upconversion nanoparticles(UCNPs) serve as NIR light-responsive nanoplatforms,showing promising applications in studying interactions between various bioactive molecules and proteins in living systems.Under NIR light irradiation,azido-naphthalimides are activated by ultraviolet(UV) and blue light emitted from UCNPs and the resulting amino-naphthalimides intermediate not only crosslink nearby target proteins but also enable imaging performance.We demonstrate that this nanoplatform is capable of selective protein labeling and imaging in complex protein environments,achieving specific labeling and imaging of both intracellular and extracellular proteins in mammalian cells as well as bacteria.Furthermore,in vivo protein labeling has been achieved using this novel NIR light-activatable nanoplatform.This technique will open new avenues for discoveries and mechanistic interrogation in chemical biology.展开更多
Cytochrome c(cyt c)is released from mitochondria into the cytosol upon apoptotic stimulation,ultimately triggering programmed cell death.Recent studies have revealed that transfer RNA(tRNA)interacts with cyt c,impedin...Cytochrome c(cyt c)is released from mitochondria into the cytosol upon apoptotic stimulation,ultimately triggering programmed cell death.Recent studies have revealed that transfer RNA(tRNA)interacts with cyt c,impeding the formation of the apoptosome complex and thereby suppressing apoptosis.To elucidate the molecular mechanism underlying the interaction between cyt c and tRNA,nuclear magnetic resonance(NMR)-based chemical shift perturbation and intensity analysis were employed to characterize the binding interface between cyt c and tRNAphe.The findings demonstrate that cyt c primarily engages with tRNAphe through its 70–85Ω-loop and N-terminalα-helix.This interaction sterically hinders the accessibility of small molecules,such as H_(2)O_(2),to the hydrophobic pocket of cyt c,consequently attenuating its peroxidase activity.Furthermore,oxidative modification of cyt c,particularly the carbonylation of positively charged lysine residues,weakens this interaction.展开更多
功能性食醋是食醋产业多元化与高值化发展的重要方向。为评估不同再加工工艺的改良效果,以“华佗药香醋”为对象,采用液相色谱-串联质谱联用(liquid chromatography-tandem mass spectrometry,LC-MS/MS)技术及体外抗氧化活性测定等方法...功能性食醋是食醋产业多元化与高值化发展的重要方向。为评估不同再加工工艺的改良效果,以“华佗药香醋”为对象,采用液相色谱-串联质谱联用(liquid chromatography-tandem mass spectrometry,LC-MS/MS)技术及体外抗氧化活性测定等方法,系统比较了浸泡(25℃)与煮沸处理对其营养成分与功能活性的影响。结果显示,浸泡工艺提升了产品的营养成分含量,总有机酸和总氨基酸分别提高了10.43%和30.04%,但未达到显著水平。同时,该工艺还显著增强了样品的抗氧化能力,1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazine,DPPH)和2,2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸[2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS]自由基清除率分别提升了87.39%和58.36%(P<0.05)。扫描电镜分析(scanning electron microscopy, SEM)进一步揭示药香醋中存在天然纳米结构,且其形貌、粒径及Zeta电位均受加工工艺调控。结果表明,浸泡工艺可同步提升药香醋的营养成分与抗氧化能力,工艺可调纳米结构为其功能增效机制提供了直接微观证据。展开更多
泛醇-细胞色素C还原酶复合物伴侣(ubiquinol-cytochrome C reductase complex chaperone,BCS1L)主要参与线粒体复合体Ⅲ组装,与线粒体遗传病发病、前列腺癌患者疲劳等有关,然而其在结肠腺癌中的作用尚不清楚。本研究通过整合多组学分析...泛醇-细胞色素C还原酶复合物伴侣(ubiquinol-cytochrome C reductase complex chaperone,BCS1L)主要参与线粒体复合体Ⅲ组装,与线粒体遗传病发病、前列腺癌患者疲劳等有关,然而其在结肠腺癌中的作用尚不清楚。本研究通过整合多组学分析和功能实验,系统探讨了BCS1L在结肠腺癌中的临床意义及分子机制。结果发现,BCS1L在结肠腺癌组织中表达水平显著上调(P<0.05),其高表达与患者不良预后及年龄显著相关(P<0.05)。功能富集结果显示,BCS1L与有机阴离子转运、羧酸转运、有机酸转运等相关信号通路有关。免疫细胞浸润结果显示,BCS1L高表达组中Treg细胞占比增多。突变图谱结果显示,PCLO、ABCA13、LRP1B、FAT3、HYDIN和SOX 9等基因突变频率在BCS1L高表达组显著升高(均P<0.05)。实验验证结果显示,敲低BCS1L能够显著抑制结肠腺癌细胞的增殖能力、克隆形成能力及迁移能力(均P<0.05)。以上研究结果表明,BCS1L可能通过调控物质转运及调节免疫抑制性肿瘤微环境在结肠腺癌细胞生长和迁移过程中发挥关键作用。展开更多
文摘综述了G蛋白偶联受体(G protein coupled receptor,GPCR)二聚化检测的生物化学、生物物理、结构生物学方法,系统阐述了相关原理与关键要点,并对比分析了优势与局限性。未来研究应聚焦于开发更加灵敏且非侵入性的GPCR二聚化检测方法,推动多种技术手段的协同联用,从而深入探索GPCR二聚体在疾病发生和发展中的作用,为药物设计提供新思路,并进一步发掘其在作物病虫害防治与畜牧业的应用潜力。
基金supported by the CJD Foundation,USA,the Alzheimer Forschung Initiative(AFI)e.V.,Germany,and Werner-Otto-Stiftung,Germany(all to HCA),ChinaScholarship Council(grant#202108080249 to FS)Deutsche Forschungsgemeinschaft(DFG)CRC877“Proteolysis as a regulatory event in pathophysiology”(project A12 to MG),Slovene Research and InnovationAgency(grant number P4-0176 to VCS).
文摘Novel insights into complex biological processes very often critically depend on the establishment of new potent read-out tools and improved protocols.A lot has been learned over the past four decades on physiological functions and,importantly,disease-related roles of the prion protein(PrP),a relatively broadly expressed membrane-anchored glycoprotein with high levels in several cell types of the nervous and immune system and with well-established key roles in different progressive and fatal neurodegenerative protein misfolding diseases(proteopathies).
基金supported by the National Natural Science Foundation of China (No.22007008)the LiaoNing Revitalization Talents Program (No.XLYC1907021)the Fundamental Research Funds for the Central Universities (Nos.DUT23YG120,DUT19RC(3)009)。
文摘The study of target proteins is crucial for understanding molecular interactions and developing analytical platforms,therapeutic agents and functional tools.Herein,we present a novel nanoplatform activated by near-infrared(NIR) light for triple-modal proteins study,which enabling target protein labeling,enrichment and visualization.Azido-naphthalimide-coated upconversion nanoparticles(UCNPs) serve as NIR light-responsive nanoplatforms,showing promising applications in studying interactions between various bioactive molecules and proteins in living systems.Under NIR light irradiation,azido-naphthalimides are activated by ultraviolet(UV) and blue light emitted from UCNPs and the resulting amino-naphthalimides intermediate not only crosslink nearby target proteins but also enable imaging performance.We demonstrate that this nanoplatform is capable of selective protein labeling and imaging in complex protein environments,achieving specific labeling and imaging of both intracellular and extracellular proteins in mammalian cells as well as bacteria.Furthermore,in vivo protein labeling has been achieved using this novel NIR light-activatable nanoplatform.This technique will open new avenues for discoveries and mechanistic interrogation in chemical biology.
基金financial support from National Key R&D Program of China(2018YFA0704002,2018YFE0202300,2023YFA1607500)National Natural Science Foundation of China(22174152,21991081,2204167,21505153,21675170,2147514621735007,and 22204167)+2 种基金Hubei Provincial Natural Science Foundation of China(2023AFA041)Strategic Priority Research Program of the Chinese Academy of Sciences(XDB0540300)Funding of Wuhan Special Project for Knowledge Innovation(2023020201010085).
文摘Cytochrome c(cyt c)is released from mitochondria into the cytosol upon apoptotic stimulation,ultimately triggering programmed cell death.Recent studies have revealed that transfer RNA(tRNA)interacts with cyt c,impeding the formation of the apoptosome complex and thereby suppressing apoptosis.To elucidate the molecular mechanism underlying the interaction between cyt c and tRNA,nuclear magnetic resonance(NMR)-based chemical shift perturbation and intensity analysis were employed to characterize the binding interface between cyt c and tRNAphe.The findings demonstrate that cyt c primarily engages with tRNAphe through its 70–85Ω-loop and N-terminalα-helix.This interaction sterically hinders the accessibility of small molecules,such as H_(2)O_(2),to the hydrophobic pocket of cyt c,consequently attenuating its peroxidase activity.Furthermore,oxidative modification of cyt c,particularly the carbonylation of positively charged lysine residues,weakens this interaction.
文摘功能性食醋是食醋产业多元化与高值化发展的重要方向。为评估不同再加工工艺的改良效果,以“华佗药香醋”为对象,采用液相色谱-串联质谱联用(liquid chromatography-tandem mass spectrometry,LC-MS/MS)技术及体外抗氧化活性测定等方法,系统比较了浸泡(25℃)与煮沸处理对其营养成分与功能活性的影响。结果显示,浸泡工艺提升了产品的营养成分含量,总有机酸和总氨基酸分别提高了10.43%和30.04%,但未达到显著水平。同时,该工艺还显著增强了样品的抗氧化能力,1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazine,DPPH)和2,2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸[2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS]自由基清除率分别提升了87.39%和58.36%(P<0.05)。扫描电镜分析(scanning electron microscopy, SEM)进一步揭示药香醋中存在天然纳米结构,且其形貌、粒径及Zeta电位均受加工工艺调控。结果表明,浸泡工艺可同步提升药香醋的营养成分与抗氧化能力,工艺可调纳米结构为其功能增效机制提供了直接微观证据。
