Despite the expansive applications of gas-phase unfolding techniques,the molecular mechanism for the solvent-free forced unfolding pathway which substrate multidomain proteins usually adopt remains elusive at the seco...Despite the expansive applications of gas-phase unfolding techniques,the molecular mechanism for the solvent-free forced unfolding pathway which substrate multidomain proteins usually adopt remains elusive at the secondary structure level.Herein,upon carefully selecting CRM_(197) as a therapeutically-relevant model system containing multiple secondary structure-separated domains,we systematically examine its solvent-free unfolding pathway.Further-more,utilizing the hybrid of noncovalent chemical probing with niacinamide and ion mobility-mass spectrometry-guided all-atom molecular dynamics simulations,we map a nearly complete unfolding atlas for the conjugate vaccine carrier protein CRM_(197) in a domain-and secondary structure-resolved manner.The totality of our data supports the preferential unfolding of the sheet-rich domain,indicating the dynamic transition from β-sheet toα-helix,and demonstrating that helix exhibit comparatively higher stability thanβ-sheets.We propose that this sheet-to-helix dynamic transition may be central to the gas-phase unfolding pathways of multidomain proteins,suggesting the need for systematic studies on additional multidomain protein systems.展开更多
The current study comprehensively evaluates four different protein extraction methods based on urea,sodium dodecyl sulfate(SDS),anionic surfactants(BT),and total RNA extractor(Trizol),aiming to optimize the sample pre...The current study comprehensively evaluates four different protein extraction methods based on urea,sodium dodecyl sulfate(SDS),anionic surfactants(BT),and total RNA extractor(Trizol),aiming to optimize the sample preparation workflow for mass spectrometry-based proteomics.Using HeLa cells as an example,we found that the method employing the mass spectrometry-compatible surfactant BT reagent significantly reduces the total time consumed for protein extraction and minimizes protein losses during the sample preparation process.Further integrating the four protein extraction methods,we identified over 7000 proteins from HeLa cells without relying on pre-fractionation techniques,and 2990 of them were quantified using label-free quantification.It is worth noting that the BT and SDS methods demonstrate higher efficiency in extracting membrane proteins,while the Urea and Trizol methods are more effective in extracting proteins from nuclear and cytoplasmic fractions.In summary,this study provides a novel solution for deep proteome coverage,particularly in the context of cellular protein extraction,by integrating mass spectrometry-compatible surfactants with traditional extraction methods to effectively enhance protein identification numbers.展开更多
Molecular aggregates are receiving tremendous attention,demonstrating immense potential for biomedical applications in vitro and in vivo.For instance,the molecular aggregates of conventional fluorophores influence the...Molecular aggregates are receiving tremendous attention,demonstrating immense potential for biomedical applications in vitro and in vivo.For instance,the molecular aggregates of conventional fluorophores influence the electronic excitation states of the aggregates,causing characteristic photophysical property changes.A fundamental understanding of this classical relationship between molecular aggregate structures and photophysics has allowed for innovative biological applications.The chemical characteristics of drug molecules generally trigger the formation of colloidal aggregates,and this is considered detrimental to the drug discovery process.Furthermore,nano-sized supramolecular aggregates have been used in biomedical imaging and therapy owing to their optimal properties for in vivo utility,including enhanced cell permeability,passive tumor targeting,and convenient surface engineering.Herein,we provide an overview of the recent trends in molecular aggregates for biomedical applications.The changes in photophysical properties of conventional fluorophores and their biological applications are discussed,followed by the effects of conventional drug molecule-aggregates on drug discovery and therapeutics development.Recent trends in the investigation of biologically important analytes with aggregation-induced emission are discussed for conventional and unconventional fluorophores.Lastly,we discuss nano-sized supramolecular aggregates used in imaging and therapeutic purposes,with a focus on in vivo utilization.展开更多
Comprehensive Summary.Atherosclerosis is a lipoprotein-driven disease.In-depth understanding of pathology and accurate identification are particularly important in clinical assessment and treatment due to the irrevers...Comprehensive Summary.Atherosclerosis is a lipoprotein-driven disease.In-depth understanding of pathology and accurate identification are particularly important in clinical assessment and treatment due to the irreversibility of atherosclerotic plaque formation.Atherosclerosis is not only accompanied by lipid droplets accumulation but also closely related to inflammation,which is accompanied by excessive reactive oxygen species(ROS)and changes in microenvironment.However,there is still a lack of a simple and rapid detection platform to simultaneously evaluate multiple indicators of atherosclerosis in multiple channels.In this study,we propose a multicolor imaging probe Cy7P-B for polarity,H_(2)O_(2) and lipid droplets to evaluate atherosclerotic plaques in vivo.Cy7P-B is sensitive to environmental polarity and can monitor polarity changes by near-infrared ratio.Moreover,Cy7P-B has H_(2)O_(2)/lipid droplets dual-analyte sequential activation characteristics.Based on the multifunctional properties of Cy7P-B,the classical biomarkers of atherosclerotic plaque,lipid accumulation and up-regulation of oxidative stress are effectively detected in atherosclerotic plaques,and more importantly,the change of aortic polarity in atherosclerosis was detected for the first time.This work provides a general molecular design approach for multi-species imaging of AS,which is helpful for effective cardiovascular disease stewardship.展开更多
Fluorescence imaging is a useful tool in the field of biomedical applications.However,its imaging capacity is limited by the depth of tissue that can be penetrated when using visible light(400-700 nm)or the first near...Fluorescence imaging is a useful tool in the field of biomedical applications.However,its imaging capacity is limited by the depth of tissue that can be penetrated when using visible light(400-700 nm)or the first near-infrared window(NIR-Ⅰ,700-900 nm).To overcome the problem,fluorescence imaging in the second near-infrared window(NIR-Ⅱ,1000-1700 nm)has been developed to reduce photon scattering,auto-absorption and tissue autofluorescence to achieve high spatiotemporal resolution and deep imaging penetration.The key to NIR-Ⅱimaging is obtaining and analyzing highly selective information from functional fluorophores that emit in the 1000-1700 nm range.With the rapid development of multidisciplinary research,various types of NIR-Ⅱfluorophores have been produced and used in non-invasive,real-time NIR-Ⅱbiomedical applications.This review summarizes some of the most prevalent NIR-Ⅱfluorophores and their synthesis,such as organic fluorophores(OFs),single-walled carbon nanotubes(SWCNTs),quantum dots(QDs),and rare-earth nanoparticles(RENPs).On this basis,we describe the applications of these fluorophores in biomedical fields,including bioimaging,biosensing,phototherapy and surgical navigation.Additionally,major challenges and prospects of NIR-Ⅱbiomedical application will be further explored.展开更多
Cell-derived microvesicles(MVs) are secreted from almost all kinds of mammalian cells into the extracellular space, and play crucial roles in intercellular communication and transporting biomolecules between cells. Ho...Cell-derived microvesicles(MVs) are secreted from almost all kinds of mammalian cells into the extracellular space, and play crucial roles in intercellular communication and transporting biomolecules between cells. However, there is a great challenge for visualizing and monitoring of MVs’ bio-behaviors due to the limitations of existing labeling methods. Herein, we report the first paradigm of designer cell-self-implemented labeling of MVs secreted from living mammalian MCF-7 cells in situ using the intracellular-synthesized fluorescent quantum dots(QDs) during the formation of MVs. By elaborately coupling intracellular biochemical reactions and metabolism pathways, the MCF-7 cells can be illuminated brightly by intracellular-biosynthesized fluorescent CdSe QDs. Simultaneously, intracellular-synthesized QDs can be in situ encapsulated by the secreted MVs budding from the plasma membrane of the fluorescing cells to label the MVs with an efficiency of up to 89.9%. The whole labeling process skillfully combines designer precise cell-tuned intricate synthesis of CdSe QDs with mild in-situ labeling via cell-selfimplementation just after feeding the cell with suitable chemicals, which is structure-or function-nondestructive and much more straightforward and milder than those by chemical conjugation or indirect encapsulation with conventional fluorogenic labels.展开更多
Photon upconversion is an anti-Stokes process that converts low-energy photons into high-energy photons.The use of upconversion luminescence can avoid the autofluorescence of biological tissue and realize background-f...Photon upconversion is an anti-Stokes process that converts low-energy photons into high-energy photons.The use of upconversion luminescence can avoid the autofluorescence of biological tissue and realize background-free bioimaging with a high signal-to-noise ratio at a low power density.In addition,the excitation of red or near-infrared light facilitates the reduction of photodamage in biological tissues and subsequent bioimaging of deep tissue features in vivo.Meanwhile,upconversion emission-mediated bio sensing offers both high sensitivity and low detection limits for quantitative analysis of the target substances in complicated biological samples.Due to its high upconversion quantum yield,low excitation power density,and tunable absorption and emission wavelengths,triplet-triplet annihilation upconversion(TTA-UC)has garnered considerable interest for bioimaging and biosensing.This review will introduce the fundamental concepts of TTA-UC,the factors that influence TTA-UC materials,and the methodologies for preparing TTA-UC materials.The important progress of TTA-UC in bioimaging and bio sensing in recent years will also be discussed in detail in vitro and in vivo.Furthermore,the current challenges of TTA-UC in bioimaging and biosensing will be discussed,along with potential solutions.展开更多
The surface of nanocrystals plays a dominant role in many of their physical and chemical properties.However,controllability and tunability of nanocrystal surfaces remain unsolved.Herein,we report that the surface chem...The surface of nanocrystals plays a dominant role in many of their physical and chemical properties.However,controllability and tunability of nanocrystal surfaces remain unsolved.Herein,we report that the surface chemistry of nanocrystals,such as near-infrared Ag_(2)Se quantum dots(QDs),is sizedependent and composition-tunable.The Ag_(2)Se QDs tend to form a stable metal complex on the surface to minimize the surface energy,and therefore the surface chemistry can be varied with particle size.Meanwhile,changes in surface inorganic composition lead to reorganization of the surface ligands,and the surface chemistry also varies with composition.Therefore,the surface chemistry of Ag_(2)Se QDs,responsible for the photoluminescence(PL)quantum yield and photostability,can be tuned by changing their size or composition.Accordingly,we demonstrate that the PL intensity of the Ag_(2)Se QDs can be tuned reversely by adjusting the degree of surface Ag^(+) enrichment via light irradiation or the addition of AgNO_(3).This work provides insight into the control of QD surface for desired PL properties.展开更多
Viral envelope fusion with the host plasma membrane(PM)for genome release is a hallmark step in the life cycle of many enveloped viruses.This process is regulated by a complex network of biomolecules on the PM,but rob...Viral envelope fusion with the host plasma membrane(PM)for genome release is a hallmark step in the life cycle of many enveloped viruses.This process is regulated by a complex network of biomolecules on the PM,but robust tools to precisely elucidate the dynamic mechanisms of virus-PM fusion events are still lacking.Here,we developed a quantitative single-virus tracking approach based on highly efficient dual-color labelling of viruses and batch trajectory analysis to achieve the spatiotemporal quantification of fusion events.This approach allows us to comprehensively analyze the membrane fusion mechanism utilized by pseudotyped severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)at the singlevirus level and precisely elucidate how the relevant biomolecules synergistically regulate the fusion process.Our results revealed that SARS-CoV-2 may promote the formation of supersaturated clusters of cholesterol to facilitate the initiation of the membrane fusion process and accelerate the viral genome release.展开更多
基金support by the National Key R&D Program of China(No.2022YFA1305200,to GL)National Natural Science Foundation of China(No.22104064 to GL,No.22173020 to JL)the US National Institute of Mental Health(No.R01MH122742,to CJW)for financial and instrumental support.
文摘Despite the expansive applications of gas-phase unfolding techniques,the molecular mechanism for the solvent-free forced unfolding pathway which substrate multidomain proteins usually adopt remains elusive at the secondary structure level.Herein,upon carefully selecting CRM_(197) as a therapeutically-relevant model system containing multiple secondary structure-separated domains,we systematically examine its solvent-free unfolding pathway.Further-more,utilizing the hybrid of noncovalent chemical probing with niacinamide and ion mobility-mass spectrometry-guided all-atom molecular dynamics simulations,we map a nearly complete unfolding atlas for the conjugate vaccine carrier protein CRM_(197) in a domain-and secondary structure-resolved manner.The totality of our data supports the preferential unfolding of the sheet-rich domain,indicating the dynamic transition from β-sheet toα-helix,and demonstrating that helix exhibit comparatively higher stability thanβ-sheets.We propose that this sheet-to-helix dynamic transition may be central to the gas-phase unfolding pathways of multidomain proteins,suggesting the need for systematic studies on additional multidomain protein systems.
文摘The current study comprehensively evaluates four different protein extraction methods based on urea,sodium dodecyl sulfate(SDS),anionic surfactants(BT),and total RNA extractor(Trizol),aiming to optimize the sample preparation workflow for mass spectrometry-based proteomics.Using HeLa cells as an example,we found that the method employing the mass spectrometry-compatible surfactant BT reagent significantly reduces the total time consumed for protein extraction and minimizes protein losses during the sample preparation process.Further integrating the four protein extraction methods,we identified over 7000 proteins from HeLa cells without relying on pre-fractionation techniques,and 2990 of them were quantified using label-free quantification.It is worth noting that the BT and SDS methods demonstrate higher efficiency in extracting membrane proteins,while the Urea and Trizol methods are more effective in extracting proteins from nuclear and cytoplasmic fractions.In summary,this study provides a novel solution for deep proteome coverage,particularly in the context of cellular protein extraction,by integrating mass spectrometry-compatible surfactants with traditional extraction methods to effectively enhance protein identification numbers.
基金Korea Institute of Science and Technology,Grant/Award Number:2E31093Korea University,Grant/Award Number:K2110571National Research Foundation of Korea,Grant/Award Numbers:2017M3A9D8029942,2018M3A9H4079286,2019M3D1A1078941,2019R1A6A1A11051471,2020R1A2C2004422,2020R1C1C1010044,2021R1A2C2005418。
文摘Molecular aggregates are receiving tremendous attention,demonstrating immense potential for biomedical applications in vitro and in vivo.For instance,the molecular aggregates of conventional fluorophores influence the electronic excitation states of the aggregates,causing characteristic photophysical property changes.A fundamental understanding of this classical relationship between molecular aggregate structures and photophysics has allowed for innovative biological applications.The chemical characteristics of drug molecules generally trigger the formation of colloidal aggregates,and this is considered detrimental to the drug discovery process.Furthermore,nano-sized supramolecular aggregates have been used in biomedical imaging and therapy owing to their optimal properties for in vivo utility,including enhanced cell permeability,passive tumor targeting,and convenient surface engineering.Herein,we provide an overview of the recent trends in molecular aggregates for biomedical applications.The changes in photophysical properties of conventional fluorophores and their biological applications are discussed,followed by the effects of conventional drug molecule-aggregates on drug discovery and therapeutics development.Recent trends in the investigation of biologically important analytes with aggregation-induced emission are discussed for conventional and unconventional fluorophores.Lastly,we discuss nano-sized supramolecular aggregates used in imaging and therapeutic purposes,with a focus on in vivo utilization.
基金the National Key Research and Development Program of China(2019YFA0210500)for H.S.Guangdong Basic and Applied Basic Research Foundation(Nos.2022A1515010133 and 2024A1515011373 to H.S.)the National Natural Science Foundation of China(No.31900033 to H.S.)forfinancial support.
文摘Comprehensive Summary.Atherosclerosis is a lipoprotein-driven disease.In-depth understanding of pathology and accurate identification are particularly important in clinical assessment and treatment due to the irreversibility of atherosclerotic plaque formation.Atherosclerosis is not only accompanied by lipid droplets accumulation but also closely related to inflammation,which is accompanied by excessive reactive oxygen species(ROS)and changes in microenvironment.However,there is still a lack of a simple and rapid detection platform to simultaneously evaluate multiple indicators of atherosclerosis in multiple channels.In this study,we propose a multicolor imaging probe Cy7P-B for polarity,H_(2)O_(2) and lipid droplets to evaluate atherosclerotic plaques in vivo.Cy7P-B is sensitive to environmental polarity and can monitor polarity changes by near-infrared ratio.Moreover,Cy7P-B has H_(2)O_(2)/lipid droplets dual-analyte sequential activation characteristics.Based on the multifunctional properties of Cy7P-B,the classical biomarkers of atherosclerotic plaque,lipid accumulation and up-regulation of oxidative stress are effectively detected in atherosclerotic plaques,and more importantly,the change of aortic polarity in atherosclerosis was detected for the first time.This work provides a general molecular design approach for multi-species imaging of AS,which is helpful for effective cardiovascular disease stewardship.
基金supported by National Key Research and Development Program of China(2019YFA0210500)National Natural Science Foundation of China(21977054,21877102 and 91953107)
文摘Fluorescence imaging is a useful tool in the field of biomedical applications.However,its imaging capacity is limited by the depth of tissue that can be penetrated when using visible light(400-700 nm)or the first near-infrared window(NIR-Ⅰ,700-900 nm).To overcome the problem,fluorescence imaging in the second near-infrared window(NIR-Ⅱ,1000-1700 nm)has been developed to reduce photon scattering,auto-absorption and tissue autofluorescence to achieve high spatiotemporal resolution and deep imaging penetration.The key to NIR-Ⅱimaging is obtaining and analyzing highly selective information from functional fluorophores that emit in the 1000-1700 nm range.With the rapid development of multidisciplinary research,various types of NIR-Ⅱfluorophores have been produced and used in non-invasive,real-time NIR-Ⅱbiomedical applications.This review summarizes some of the most prevalent NIR-Ⅱfluorophores and their synthesis,such as organic fluorophores(OFs),single-walled carbon nanotubes(SWCNTs),quantum dots(QDs),and rare-earth nanoparticles(RENPs).On this basis,we describe the applications of these fluorophores in biomedical fields,including bioimaging,biosensing,phototherapy and surgical navigation.Additionally,major challenges and prospects of NIR-Ⅱbiomedical application will be further explored.
基金the National Natural Science Foundation of China(21535005,91859123,21705111).
文摘Cell-derived microvesicles(MVs) are secreted from almost all kinds of mammalian cells into the extracellular space, and play crucial roles in intercellular communication and transporting biomolecules between cells. However, there is a great challenge for visualizing and monitoring of MVs’ bio-behaviors due to the limitations of existing labeling methods. Herein, we report the first paradigm of designer cell-self-implemented labeling of MVs secreted from living mammalian MCF-7 cells in situ using the intracellular-synthesized fluorescent quantum dots(QDs) during the formation of MVs. By elaborately coupling intracellular biochemical reactions and metabolism pathways, the MCF-7 cells can be illuminated brightly by intracellular-biosynthesized fluorescent CdSe QDs. Simultaneously, intracellular-synthesized QDs can be in situ encapsulated by the secreted MVs budding from the plasma membrane of the fluorescing cells to label the MVs with an efficiency of up to 89.9%. The whole labeling process skillfully combines designer precise cell-tuned intricate synthesis of CdSe QDs with mild in-situ labeling via cell-selfimplementation just after feeding the cell with suitable chemicals, which is structure-or function-nondestructive and much more straightforward and milder than those by chemical conjugation or indirect encapsulation with conventional fluorogenic labels.
基金the financial support provided by Research start-up fund of Nankai UniversityTianjin Natural Science Foundation(S22QNG922)+1 种基金National Natural Science Fund for Excellent Young Scientists Fund Program(Overseas)(013398)the Open Fund of the State Key Laboratory of Fine Chemicals(Dalian University of Technology)(KF2111)
文摘Photon upconversion is an anti-Stokes process that converts low-energy photons into high-energy photons.The use of upconversion luminescence can avoid the autofluorescence of biological tissue and realize background-free bioimaging with a high signal-to-noise ratio at a low power density.In addition,the excitation of red or near-infrared light facilitates the reduction of photodamage in biological tissues and subsequent bioimaging of deep tissue features in vivo.Meanwhile,upconversion emission-mediated bio sensing offers both high sensitivity and low detection limits for quantitative analysis of the target substances in complicated biological samples.Due to its high upconversion quantum yield,low excitation power density,and tunable absorption and emission wavelengths,triplet-triplet annihilation upconversion(TTA-UC)has garnered considerable interest for bioimaging and biosensing.This review will introduce the fundamental concepts of TTA-UC,the factors that influence TTA-UC materials,and the methodologies for preparing TTA-UC materials.The important progress of TTA-UC in bioimaging and bio sensing in recent years will also be discussed in detail in vitro and in vivo.Furthermore,the current challenges of TTA-UC in bioimaging and biosensing will be discussed,along with potential solutions.
基金supported by the National Natural Science Foundation of China(91859123)the National Key R&D Program of China(2019YFA0210103)。
文摘The surface of nanocrystals plays a dominant role in many of their physical and chemical properties.However,controllability and tunability of nanocrystal surfaces remain unsolved.Herein,we report that the surface chemistry of nanocrystals,such as near-infrared Ag_(2)Se quantum dots(QDs),is sizedependent and composition-tunable.The Ag_(2)Se QDs tend to form a stable metal complex on the surface to minimize the surface energy,and therefore the surface chemistry can be varied with particle size.Meanwhile,changes in surface inorganic composition lead to reorganization of the surface ligands,and the surface chemistry also varies with composition.Therefore,the surface chemistry of Ag_(2)Se QDs,responsible for the photoluminescence(PL)quantum yield and photostability,can be tuned by changing their size or composition.Accordingly,we demonstrate that the PL intensity of the Ag_(2)Se QDs can be tuned reversely by adjusting the degree of surface Ag^(+) enrichment via light irradiation or the addition of AgNO_(3).This work provides insight into the control of QD surface for desired PL properties.
基金supported by the National Natural Science Foundation of China(22293032,22293030,and 91859123)the National Key Research and Development Program of China(2019YFA0210500)+1 种基金the Fundamental Research Funds for the Central Universities of China(63211023)the financial support from Haihe Laboratory of Sustainable Chemical Transformations.
文摘Viral envelope fusion with the host plasma membrane(PM)for genome release is a hallmark step in the life cycle of many enveloped viruses.This process is regulated by a complex network of biomolecules on the PM,but robust tools to precisely elucidate the dynamic mechanisms of virus-PM fusion events are still lacking.Here,we developed a quantitative single-virus tracking approach based on highly efficient dual-color labelling of viruses and batch trajectory analysis to achieve the spatiotemporal quantification of fusion events.This approach allows us to comprehensively analyze the membrane fusion mechanism utilized by pseudotyped severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)at the singlevirus level and precisely elucidate how the relevant biomolecules synergistically regulate the fusion process.Our results revealed that SARS-CoV-2 may promote the formation of supersaturated clusters of cholesterol to facilitate the initiation of the membrane fusion process and accelerate the viral genome release.
