Recently,I attended a conference organised by the European Food Safety Authority in the beautiful and prosperous Italian city of Parma.The overall topic of the conference was risk assessment,and the program included a...Recently,I attended a conference organised by the European Food Safety Authority in the beautiful and prosperous Italian city of Parma.The overall topic of the conference was risk assessment,and the program included a section on aspects of environmental risk assessment.In various areas,including the evalution of the effects of pesticide applications,invasive organisms or genetically modified plants(Arpaia et al.,2014)preparing an environmental risk assessment is an obviously relevant exercise.展开更多
Hydroxyl-terminated polybutadiene/toluene diisocyanate(HTPB/TDI)system is widely used in composite solid propellants.The migrations of plasticizers and water molecules from solid propellants and surrounding environmen...Hydroxyl-terminated polybutadiene/toluene diisocyanate(HTPB/TDI)system is widely used in composite solid propellants.The migrations of plasticizers and water molecules from solid propellants and surrounding environment to the inhibitor have always been the important issues.This study focuses on the preparation,characterization and anti-migration behavior of graphene oxide(GO)/HTPB nanocomposite liner.The GO/HTPB(GH)composite liners affect the migration of small molecules through a tighter cross-linked structure and weakening function of small molecule adsorption.The anti-migration performance of the liner at different temperatures was analyzed,and the influence of the added amount of GO on the anti-migration performance and adhesion performance was also systematically studied.The overall performance of the liner is optimized when the amount of GO filler is 0.3 wt%.After adding 0.3 wt%GO,the concentration of dioctyl sebacate(DOS)migrated into the liner is decreased by 23.28%,and the concentration of water molecules is decreased by 51.89%,indicating that the introduction of GO can significantly improve the anti-migration performance of the liner.In addition,the bond strength is greatly increased from 0.25 MPa to 0.95 MPa,which meets the application requirements of the current propellant system.This research provides an important way for the preparation of structure-function synergistic anti-migration composite liners.展开更多
Von Willebrand Factor(VWF),a multimeric plasma glycoprotein,is synthesized in endothelial cells and megakaryocytes.In adhesion and aggregation of circulating platelets towards to the sites of vascular injury,VWF captu...Von Willebrand Factor(VWF),a multimeric plasma glycoprotein,is synthesized in endothelial cells and megakaryocytes.In adhesion and aggregation of circulating platelets towards to the sites of vascular injury,VWF captures and activates the circulating platelets through interaction with platelet GPlba.As a triplet complex of A1A2A3,the VWF-A domain is a closed conformation with a low affinity to GPlba,but mutations or pathological hemodynamic environment of high fluid shear stress can induce the closed A domain to become an extended one.However,the key events in the force-and/or mutation-induced activation of VWF-A under flows remains unclear.Therefore,with techniques of AFM and PPFC,we here examined transformation of conformation and function of VWF-A under various wall shear stresses,for understanding regulation of force on VWF-A activation.Interesting,AFM scanning imaging data showed that VWF-A molecules on substrate pretreated by perfusing distilled water at various wall shear stresses shortened first and then lengthened as increasing of the pre-loaded wall shear stress,and the threshold of the wall shear stress is about 100 dyn/cm2,demonstrating that increasing pre-loaded wall shear stress would make the treated-A1A2A3 conformation gradually transform from a loose spherical structure to a compact one first and then become an open or extended one.The adhesion frequency of GPlba-coated Polystyrene microspheres(3-μm radius)on the VWF-A-coated substrates decreased first and then increased with the preloaded wall shear stress,which has a same threshold mentioned above.These results suggested that,force-induced activation of VWF-A occurs just at high wall shear stresses(>100 dyn/cm2).The mechanical stability of the closed A1A2A3 conformation would be weakened by the gain of function(GOF)mutant R1 308 L of A1 and enhanced by the loss of function(LOF)mutant G1324S,as it should be.To further reveal the molecular mechanism of the force-induced enhancing or weakening of VWF-A activation,we performed AFM experiment to investigate interaction of A1(WT A1 and its two mutants,the GOF mutant R1 308 L and the LOF mutant G1324S)with A2 and A3,respectively.The adhesive frequency of A1 with A2was larger than that of A1 with A3,showing that A1 was in favor of A2 rather than A3.And,the lifetimes of A1-A2 and A1-A3bond were biphasic force-dependent,showing a'Catch-slip bond'transform in binding of A1 to A2 or A3.It suggested that under the low wall shear stresses,force could inhibit VWF activity through a catch bond mechanism,which enhanced the stability of the closed A1A2A3 conformation,but under high wall shear stresses,the force would enhance VWF activity through a slip bond mechanism,which promoted conformational transform of VWF-A from closed to extended one through reducing the stability of the closed A1A2A3 structure.Our results showed that the GOF mutant R1 308 L would down-regulate the binding affinity of A1 to A2,leading to a low barrier in opening of the closed VWF-A structure.In contrast,the LOF mutant G1324S would enhance the stability of the closed VWF-A conformation by up-regulating the binding affinity of A1 to A2,leading to inhibition of VWF activity.展开更多
Integrin activation,the transition from a low to a high affinity state,regulates the numerous cellular responses consequent to integrin engagement by extracellular matrix proteins.Kindlin proteins,play crucial roles i...Integrin activation,the transition from a low to a high affinity state,regulates the numerous cellular responses consequent to integrin engagement by extracellular matrix proteins.Kindlin proteins,play crucial roles in the integrin-signaling pathway by directly interacting with and activating integrins,which mediate the cell-extracellular matrix adhesion and signaling.As a widely distributed PTB domain protein and a major member of the kindlin family,kindlin2 interacts withβ3-tail,bridges talin-activated integrins to promote integrin aggregation,and enhances talin-induced integrin activation.Thus,kindlin2 is identified as a coactivator of integrins.Unlike talins,kindlin2 cannot directly alter the conformation of the integrin transmembrane helix and fail to activate integrin alone.Nevertheless,although it is widely accepted that kindlins and talins synergistically promote integrin activation,the underlying mechanism is unclear.Thus,the study of the force dissociation of the kindlin2/β3-tail complex and the conformation stabilization under different mechanical micro-environments should be of great significance for the further understanding of the structural basis of its synergistically activation of integrin.To reveal the molecular dynamics mechanism of interaction between kindlin2 andβ3-tail,we perform molecular dynamics(MD)simulations for this complex with different computing strategies interaction.In MD simulations,the available crystal structures of Kindlin-2/β3-tail complex(Protein Data Bank code 5XQ1)was downloaded from the PDB database.Two software packages,VMD for visualization and modeling and NAMD 2.13 for energy minimizations and MD simulations,were used here.The steadystate conformation of the complex was obtained from the equilibrium simulation.The dissociation event was observed by the constant velocity simulation,and the mechanical stability of the complex was observed by the constant force simulation.Our results showed that,during the equilibrium of the kindlin2-F3/β34ail complex,the residue MET612,LYS613 and TRP615 on the F3 domain of kindlin2 contributed to hydrogen-bonding with the corresponding residues onβ3 integrin.These bonds exhibit moderate or strong stability through steered molecular dynamics(SMD)simulation.During the constant velocity simulation,the complex exhibits a variety of unfolding pathways against tension applications,which are mainly distinguished by the disruption of hydrogen-bonds between the F3 domain a1/a2 helixes andβ1/β2 sheets.During the constant force simulation,the different phases of the composite force dissociation have different dissociation probabilities,which shows the biphasic force-dependent characteristics.And,the key residues in the pulling were recognized according not only to the number of interacting residue pairs,but also to their bond strength.Using molecular dynamics simulation,we showed the steady state of the kindlin2-F3/β3-tail complex under different tensile forces,and observe the dynamic process of molecular interaction.A possible underlying biophysical mechanism is that,the dissociation of Kindlin2-F3/β3-tail complex is biphasic force-dependent,and the conformations under different stretching states have different binding affinities.This study not only provides insights into the structural basis and mechanical regulation mechanisms of the kindlin/integrin interaction,in understanding in kindlin/integrin-related signaling in different cellular biological processes,but also provides new ideas for novel drug design and the treatment of related diseases.展开更多
文摘Recently,I attended a conference organised by the European Food Safety Authority in the beautiful and prosperous Italian city of Parma.The overall topic of the conference was risk assessment,and the program included a section on aspects of environmental risk assessment.In various areas,including the evalution of the effects of pesticide applications,invasive organisms or genetically modified plants(Arpaia et al.,2014)preparing an environmental risk assessment is an obviously relevant exercise.
基金the financial support of the National Natural Science Foundation of China(grant number 22005145)the Natural Science Foundation of Jiangsu Province(grant number BK20180495,BK20180698)+1 种基金the Opening Project of Key Laboratory of Special Energy Materials(Nanjing University of Science and Technology)the Fundamental Research Funds for the Priority Academic Program Development of Jiangsu Higher Education Institutions(grant number 30919011404)。
文摘Hydroxyl-terminated polybutadiene/toluene diisocyanate(HTPB/TDI)system is widely used in composite solid propellants.The migrations of plasticizers and water molecules from solid propellants and surrounding environment to the inhibitor have always been the important issues.This study focuses on the preparation,characterization and anti-migration behavior of graphene oxide(GO)/HTPB nanocomposite liner.The GO/HTPB(GH)composite liners affect the migration of small molecules through a tighter cross-linked structure and weakening function of small molecule adsorption.The anti-migration performance of the liner at different temperatures was analyzed,and the influence of the added amount of GO on the anti-migration performance and adhesion performance was also systematically studied.The overall performance of the liner is optimized when the amount of GO filler is 0.3 wt%.After adding 0.3 wt%GO,the concentration of dioctyl sebacate(DOS)migrated into the liner is decreased by 23.28%,and the concentration of water molecules is decreased by 51.89%,indicating that the introduction of GO can significantly improve the anti-migration performance of the liner.In addition,the bond strength is greatly increased from 0.25 MPa to 0.95 MPa,which meets the application requirements of the current propellant system.This research provides an important way for the preparation of structure-function synergistic anti-migration composite liners.
基金supported by the National Natural Science Foundation of China ( 116272109,11432006)
文摘Von Willebrand Factor(VWF),a multimeric plasma glycoprotein,is synthesized in endothelial cells and megakaryocytes.In adhesion and aggregation of circulating platelets towards to the sites of vascular injury,VWF captures and activates the circulating platelets through interaction with platelet GPlba.As a triplet complex of A1A2A3,the VWF-A domain is a closed conformation with a low affinity to GPlba,but mutations or pathological hemodynamic environment of high fluid shear stress can induce the closed A domain to become an extended one.However,the key events in the force-and/or mutation-induced activation of VWF-A under flows remains unclear.Therefore,with techniques of AFM and PPFC,we here examined transformation of conformation and function of VWF-A under various wall shear stresses,for understanding regulation of force on VWF-A activation.Interesting,AFM scanning imaging data showed that VWF-A molecules on substrate pretreated by perfusing distilled water at various wall shear stresses shortened first and then lengthened as increasing of the pre-loaded wall shear stress,and the threshold of the wall shear stress is about 100 dyn/cm2,demonstrating that increasing pre-loaded wall shear stress would make the treated-A1A2A3 conformation gradually transform from a loose spherical structure to a compact one first and then become an open or extended one.The adhesion frequency of GPlba-coated Polystyrene microspheres(3-μm radius)on the VWF-A-coated substrates decreased first and then increased with the preloaded wall shear stress,which has a same threshold mentioned above.These results suggested that,force-induced activation of VWF-A occurs just at high wall shear stresses(>100 dyn/cm2).The mechanical stability of the closed A1A2A3 conformation would be weakened by the gain of function(GOF)mutant R1 308 L of A1 and enhanced by the loss of function(LOF)mutant G1324S,as it should be.To further reveal the molecular mechanism of the force-induced enhancing or weakening of VWF-A activation,we performed AFM experiment to investigate interaction of A1(WT A1 and its two mutants,the GOF mutant R1 308 L and the LOF mutant G1324S)with A2 and A3,respectively.The adhesive frequency of A1 with A2was larger than that of A1 with A3,showing that A1 was in favor of A2 rather than A3.And,the lifetimes of A1-A2 and A1-A3bond were biphasic force-dependent,showing a'Catch-slip bond'transform in binding of A1 to A2 or A3.It suggested that under the low wall shear stresses,force could inhibit VWF activity through a catch bond mechanism,which enhanced the stability of the closed A1A2A3 conformation,but under high wall shear stresses,the force would enhance VWF activity through a slip bond mechanism,which promoted conformational transform of VWF-A from closed to extended one through reducing the stability of the closed A1A2A3 structure.Our results showed that the GOF mutant R1 308 L would down-regulate the binding affinity of A1 to A2,leading to a low barrier in opening of the closed VWF-A structure.In contrast,the LOF mutant G1324S would enhance the stability of the closed VWF-A conformation by up-regulating the binding affinity of A1 to A2,leading to inhibition of VWF activity.
基金supported by the National Natural Science Foundation of China ( 116272109, 11432006)
文摘Integrin activation,the transition from a low to a high affinity state,regulates the numerous cellular responses consequent to integrin engagement by extracellular matrix proteins.Kindlin proteins,play crucial roles in the integrin-signaling pathway by directly interacting with and activating integrins,which mediate the cell-extracellular matrix adhesion and signaling.As a widely distributed PTB domain protein and a major member of the kindlin family,kindlin2 interacts withβ3-tail,bridges talin-activated integrins to promote integrin aggregation,and enhances talin-induced integrin activation.Thus,kindlin2 is identified as a coactivator of integrins.Unlike talins,kindlin2 cannot directly alter the conformation of the integrin transmembrane helix and fail to activate integrin alone.Nevertheless,although it is widely accepted that kindlins and talins synergistically promote integrin activation,the underlying mechanism is unclear.Thus,the study of the force dissociation of the kindlin2/β3-tail complex and the conformation stabilization under different mechanical micro-environments should be of great significance for the further understanding of the structural basis of its synergistically activation of integrin.To reveal the molecular dynamics mechanism of interaction between kindlin2 andβ3-tail,we perform molecular dynamics(MD)simulations for this complex with different computing strategies interaction.In MD simulations,the available crystal structures of Kindlin-2/β3-tail complex(Protein Data Bank code 5XQ1)was downloaded from the PDB database.Two software packages,VMD for visualization and modeling and NAMD 2.13 for energy minimizations and MD simulations,were used here.The steadystate conformation of the complex was obtained from the equilibrium simulation.The dissociation event was observed by the constant velocity simulation,and the mechanical stability of the complex was observed by the constant force simulation.Our results showed that,during the equilibrium of the kindlin2-F3/β34ail complex,the residue MET612,LYS613 and TRP615 on the F3 domain of kindlin2 contributed to hydrogen-bonding with the corresponding residues onβ3 integrin.These bonds exhibit moderate or strong stability through steered molecular dynamics(SMD)simulation.During the constant velocity simulation,the complex exhibits a variety of unfolding pathways against tension applications,which are mainly distinguished by the disruption of hydrogen-bonds between the F3 domain a1/a2 helixes andβ1/β2 sheets.During the constant force simulation,the different phases of the composite force dissociation have different dissociation probabilities,which shows the biphasic force-dependent characteristics.And,the key residues in the pulling were recognized according not only to the number of interacting residue pairs,but also to their bond strength.Using molecular dynamics simulation,we showed the steady state of the kindlin2-F3/β3-tail complex under different tensile forces,and observe the dynamic process of molecular interaction.A possible underlying biophysical mechanism is that,the dissociation of Kindlin2-F3/β3-tail complex is biphasic force-dependent,and the conformations under different stretching states have different binding affinities.This study not only provides insights into the structural basis and mechanical regulation mechanisms of the kindlin/integrin interaction,in understanding in kindlin/integrin-related signaling in different cellular biological processes,but also provides new ideas for novel drug design and the treatment of related diseases.
