Metabolic dysfunction-associated steatotic liver disease(MASLD)is a metabolic disease that can progress to metabolic dysfunction-associated steatohepatitis(MASH),cirrhosis,and cancer.The zonal distribution of biomolec...Metabolic dysfunction-associated steatotic liver disease(MASLD)is a metabolic disease that can progress to metabolic dysfunction-associated steatohepatitis(MASH),cirrhosis,and cancer.The zonal distribution of biomolecules in the liver is implicated in mediat-ing the disease progression.Recently,G-protein-coupled receptor 35(GPR35)has been highlighted to play a role in MASLD,but the precise mechanism is not fully understood,particularly,in a liver-zonal manner.Here,we aimed to identify spatially distributed specific genes and metabolites in different liver zonation that are regulated by GPR35 in MASLD,by combining lipid metabolomics,spatial transcriptomics(ST),and spatial metabolomics(SM).We found that GPR35 influenced lipid accumulation,inflammatory and metabolism-related factors in specific regions,notably affecting the anti-inflammation factor ELF4(E74 like E-twenty six(ETS)tran-scription factor 4),lipid homeostasis key factor CIDEA(cell death-inducing DNA fragmentation factor alpha(DFFA)-like effector A),and the injury response-related genes SAA1/2/3(serum amyloid A1/2/3),thereby impacting MASLD progression.Furthermore,SM elucidated specific metabolite distributions across different liver regions,such as C10H11N4O7P(3ʹ,5ʹ-cyclic inosine monophosphate(3ʹ,5ʹ-IMP))for the central vein,and this metabolite significantly decreased in the liver zones of GPR35-deficient mice during MASLD progression.Taken together,GPR35 regulates hepatocyte damage repair,controls inflammation,and prevents MASLD progression by influencing phospholipid homeostasis and gene expression in a zonal manner.展开更多
Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),is characterized by a strong production of inflammatory cytokines such as TNF and IL-6,which underlie the severi...Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),is characterized by a strong production of inflammatory cytokines such as TNF and IL-6,which underlie the severity of the disease.However,the molecular mechanisms responsible for such a strong immune response remains unclear.Here,utilizing targeted tandem mass spectrometry to analyze serum metabolome and lipidome in COVID-19 patients at different temporal stages,we identified that 611 metabolites(of 1,039)were significantly altered in COVID-19 patients.Among them,two metabolites,agmatine and putrescine,were prominently elevated in the serum of patients;and 2-quinolinecarboxylate was changed in a biphasic manner,elevated during early COVID-19 infection but levelled off.When tested in mouse embryonic fibroblasts(MEFs)and macrophages,these 3 metabolites were found to activate the NF-κB pathway that plays a pivotal role in governing cytokine production.Importantly,these metabolites were each able to cause strong increase of TNF and IL-6 levels when administered to wildtype mice,but not in the mice lacking NF-κB.Intriguingly,these metabolites have little effects on the activation of interferon regulatory factors(IRFs)for the production of type I interferons(IFNs)for antiviral defenses.These data suggest that circulating metabolites resulting from COVID-19 infection may act as effectors to elicit the peculiar systemic inflammatory responses,exhibiting severely strong proinflammatory cytokine production with limited induction of the interferons.Our study may provide a rationale for development of drugs to alleviate inflammation in COVID-19 patients.展开更多
Background:Y-box binding protein 1(YB1 or YBX1)plays a critical role in tumorigenesis and cancer progression.However,whether YB1 affects malignant transformation by modulating non-codingRNAs remains largely unknown.Th...Background:Y-box binding protein 1(YB1 or YBX1)plays a critical role in tumorigenesis and cancer progression.However,whether YB1 affects malignant transformation by modulating non-codingRNAs remains largely unknown.This study aimed to investigate the relationship between YB1 and microRNAs and reveal the underlying mechanism by which YB1 impacts on tumor malignancy via miRNAs-mediated regulatory network.Methods:The biological functions of YB1 in hepatocellular carcinoma(HCC)cells were investigated by cell proliferation,wound healing,and transwell invasion assays.The miRNAs dysregulated by YB1 were screened by microarray analysis in HCC cell lines.The regulation of YB1 on miR-205 and miR-200b was determined by quantitative real-time PCR,dual-luciferase reporter assay,RNA immunoprecipitation,and pull-down assay.The relationships of YB1,DGCR8,Dicer,TUT4,and TUT1 were identified by pull-down and coimmunoprecipitation experiments.The cellular co-localization of YB1,DGCR8,and Dicer were detected by immunofluorescent staining.The in vivo effect of YB1 on tumor metastasis was determined by injecting MHCC97H cells transduced with YB1 shRNA or shControl via the tail vein in nude BALB/c mice.The expression levels of epithelial tomesenchymal transition markerswere detected by immunoblotting and immunohistochemistry assays.Results:YB1 promoted HCC cell migration and tumor metastasis by regulating miR-205/200b‒ZEB1 axis partially in a Snail-independent manner.YB1 suppressedmiR-205 and miR-200b maturation by interacting with the microprocessors DGCR8 and Dicer as well as TUT4 and TUT1 via the conserved cold shock domain.Subsequently,the downregulation of miR-205 and miR-200b enhanced ZEB1 expression,thus leading to increased cell migration and invasion.Furthermore,statistical analyses on gene expression data from HCC and normal liver tissues showed that YB1 expression was positively associated with ZEB1 expression and remarkably correlated with clinical prognosis.Conclusion:This study reveals a previously undescribed mechanism by which YB1 promotes cancer progression by regulating the miR-205/200b‒ZEB1 axis in HCC cells.Furthermore,these results highlight that YB1 may play biological functions via miRNAs-mediated gene regulation,and it can serve as a potential therapeutic target in human cancers.展开更多
Mammalian target of rapamycin(mTOR)controls cellular anabolism,and mTOR signaling is hyperactive in most cancer cells.As a result,inhibition of mTOR signaling benefits cancer patients.Rapamycin is a US Food and Drug A...Mammalian target of rapamycin(mTOR)controls cellular anabolism,and mTOR signaling is hyperactive in most cancer cells.As a result,inhibition of mTOR signaling benefits cancer patients.Rapamycin is a US Food and Drug Administration(FDA)-approved drug,a specific mTOR complex 1(mTORC1)inhibitor,for the treatment of several different types of cancer.However,rapamycin is reported to inhibit cancer growth rather than induce apoptosis.Pyruvate dehydrogenase complex(PDHc)is the gatekeeper for mitochondrial pyruvate oxidation.PDHc inactivation has been observed in a number of cancer cells,and this alteration protects cancer cells from senescence and nicotinamide adenine dinucleotide(NAD^(+))exhaustion.In this paper,we describe our finding that rapamycin treatment promotes pyruvate dehydrogenase E1 subunit alpha 1(PDHA1)phosphorylation and leads to PDHc inactivation dependent on mTOR signaling inhibition in cells.This inactivation reduces the sensitivity of cancer cells'response to rapamycin.As a result,rebooting PDHc activity with dichloroacetic acid(DCA),a pyruvate dehydrogenase kinase(PDK)inhibitor,promotes cancer cells'susceptibility to rapamycin treatment in vitro and in vivo.展开更多
Protein-metabolite interactions(PMIs)play important roles in various biological processes,especially in disease progression.However,due to the complexity of living cells,it is very difficult to identify specific PMIs....Protein-metabolite interactions(PMIs)play important roles in various biological processes,especially in disease progression.However,due to the complexity of living cells,it is very difficult to identify specific PMIs.Herein,we chose one oncogenic factor,metadherin(MTDH),as a bait to identify its in vivo interacting metabolites in cancer cells.Cholesterol is an important metabolite and essential structural component of cell membranes.It could also drive several diseases including cancer.Interestingly,we found that cholesterol robustly interacted with MTDH and downregulated the expression of MTDH in cancer cells.Furthermore,MTDH disturbed metabolite alterations under cholesterol treatment in MTDH transduced cancer cells.Collectively,our results uncover an undescribed PMI where MTDH,as an oncogenic factor,might positively regulate cancer progression by interacting with choleste rol.This study interprets the theoretical basis of PMI-oriented cancer progression and targeting therapies in clinic.展开更多
Pyruvate is an essential fuel for maintaining the tricarboxylic acid(TCA)cycle in the mitochondria.However,the precise mole-cular mechanism of pyruvate uptake by mitochondrial pyruvate carrier(MPC)is largely unknown.H...Pyruvate is an essential fuel for maintaining the tricarboxylic acid(TCA)cycle in the mitochondria.However,the precise mole-cular mechanism of pyruvate uptake by mitochondrial pyruvate carrier(MPC)is largely unknown.Here,we report that the DNA/RNA-binding protein Y-box binding protein 1(YBX1)is localized to the mitochondrial inter-membrane space by its C-terminal domain(CTD)in cancer cells.In mitochondria,YBX1 inhibits pyruvate uptake by associating with MPC1/2,thereby suppressing pyruvate-dependent TCA cycle flux.This association,in turn,promotes MPC-mediated glutaminolysis and histone lactylation.Our findings reveal that the YBX1-MPC axis exhibits a positive correlation with metastatic potential,while does not affect cell proliferation in both cultured cells and tumor xenografts.Therefore,the restricted pyruvate uptake into mitochondria potentially represents a hallmark of metastatic capacity,suggesting that the YBX1-MPC axis is a therapeutic target for combating cancer metastasis.展开更多
Glutathione S-transferases(GSTs),detoxification enzymes that catalyze the addition of glutathione(GSH)to diverse electrophilic molecules,are often overexpressed in various tumor cells.While fluorescent probes for GSTs...Glutathione S-transferases(GSTs),detoxification enzymes that catalyze the addition of glutathione(GSH)to diverse electrophilic molecules,are often overexpressed in various tumor cells.While fluorescent probes for GSTs have often adopted the 2,4-dinitrobenzenesulfonyl(DNs)group as the receptor unit,they usually suffer from considerable background reaction noise with GSH due to excessive electron deficiency.However,weakening this reactivity is generally accompanied by loss of sensitivity for GSTs,and therefore,finely turning down the reactivity while maintaining certain sensitivity is critical for developing a practical probe.Here,we report a rational semiquantitative strategy for designing such a practical two-photon probe by introducing a parameter adopted from the conceptual density functional theory(CDFT),the local electrophilicityω_(k),to characterize this reactivity.As expected,kinetic studies establishedω_(k)as efficient to predict the reactivity with GSH,and probe NI3 showing the best performance was successfully applied to detecting GST activities in live cells and tissue sections with high sensitivity and signal-to-noise ratio.Photoinduced electron transfer of naphthalimide-based probes,captured by femtosecond transient absorption for the first time and unraveled by theoretical calculations,also contributes to the negligible background noise.展开更多
Active endogenous metabolites regulate the viability of cells. This process is controlled by a series ofinteractions between small metabolites and large proteins. Previously, several studies had reported thatmetabolit...Active endogenous metabolites regulate the viability of cells. This process is controlled by a series ofinteractions between small metabolites and large proteins. Previously, several studies had reported thatmetabolite regulates the protein functions, such as diacylglycerol to protein kinase C, lactose regulationof the lac repressor, and HIF-1α stabilization by 2-hydroxyglutarate. However, decades old traditionalbiochemical methods are insufficient to systematically investigate the bio-molecular reactions for a high-throughput discovery. Here, we have reviewed an update on the recently developed chemical proteomicscalled activity-based protein profiling （ABPP）. ABPP is able to identify proteins interacted eithercovalently or non-covalently with metabolites significantly. Thus, ABPP will facilitate the characteriza-tion of specific metabolite regulating; proteins in human disease progression.展开更多
Metabolites can double as a signaling modality that initiates physiological adaptations.Metabolism,a chemical language encoding biological information,has been recognized as a powerful principle directing inflammatory...Metabolites can double as a signaling modality that initiates physiological adaptations.Metabolism,a chemical language encoding biological information,has been recognized as a powerful principle directing inflammatory responses.Cytosolic pH is a regulator of inflammatory response in macrophages.Here,we found that L-malate exerts anti-inflammatory effect via BiP-IRF2BP2 signaling,which is a sensor of cytosolic pH in macrophages.First,L-malate,a TCA intermediate upregulated in pro-inflammatory macrophages,was identified as a potent anti-inflammatory metabolite through initial screening.Subsequent screening with DARTS and MS led to the isolation of L-malate-BiP binding.Further screening through protein‒protein interaction microarrays identified a L-malate-restrained coupling of BiP with IRF2BP2,a known anti-inflammatory protein.Interestingly,pH reduction,which promotes carboxyl protonation of L-malate,facilitates L-malate and carboxylate analogues such as succinate to bind BiP,and disrupt BiPIRF2BP2 interaction in a carboxyl-dependent manner.Both L-malate and acidification inhibit BiP-IRF2BP2 interaction,and protect IRF2BP2 from BiP-driven degradation in macrophages.Furthermore,both in vitro and in vivo,BiP-IRF2BP2 signal is required for effects of both L-malate and pH on inflammatory responses.Thesefindings reveal a previously unrecognized,proton/carboxylate dual sensing pathway wherein pH and L-malate regulate inflammatory responses,indicating the role of certain carboxylate metabolites as adaptors in the proton biosensing by interactions between macromolecules.展开更多
基金supported by the National Key Research and Development Program of China(2022YFA0806503)the National Natural Science Foundation of China(81972625 and 32201217)+3 种基金Liaoning Revitalization Talents Program(XLYC2002035)Liaoning Science and Technology Innovation Funding(20230101-JH2/1013)the Innovation Program of Science and Research from Dalian Institute of Chemical Physics,Chinese Academy of Sciences(DICP I202129 and DICP I202209)the Science and Technology Innovation Fund(Youth Science and Technology Star)of Dalian(2021RQ009 and 2023RQ040).
文摘Metabolic dysfunction-associated steatotic liver disease(MASLD)is a metabolic disease that can progress to metabolic dysfunction-associated steatohepatitis(MASH),cirrhosis,and cancer.The zonal distribution of biomolecules in the liver is implicated in mediat-ing the disease progression.Recently,G-protein-coupled receptor 35(GPR35)has been highlighted to play a role in MASLD,but the precise mechanism is not fully understood,particularly,in a liver-zonal manner.Here,we aimed to identify spatially distributed specific genes and metabolites in different liver zonation that are regulated by GPR35 in MASLD,by combining lipid metabolomics,spatial transcriptomics(ST),and spatial metabolomics(SM).We found that GPR35 influenced lipid accumulation,inflammatory and metabolism-related factors in specific regions,notably affecting the anti-inflammation factor ELF4(E74 like E-twenty six(ETS)tran-scription factor 4),lipid homeostasis key factor CIDEA(cell death-inducing DNA fragmentation factor alpha(DFFA)-like effector A),and the injury response-related genes SAA1/2/3(serum amyloid A1/2/3),thereby impacting MASLD progression.Furthermore,SM elucidated specific metabolite distributions across different liver regions,such as C10H11N4O7P(3ʹ,5ʹ-cyclic inosine monophosphate(3ʹ,5ʹ-IMP))for the central vein,and this metabolite significantly decreased in the liver zones of GPR35-deficient mice during MASLD progression.Taken together,GPR35 regulates hepatocyte damage repair,controls inflammation,and prevents MASLD progression by influencing phospholipid homeostasis and gene expression in a zonal manner.
基金supported by the National Natural Science Foundation of China(31922034,82088102,91854208,92057204,31730058,82072777)Science and Technology Program of Fujian Provincial Health Commission(2021ZD02006)+3 种基金Xiamen Science and Technology Major Project(3502Z2020YJ05)Xiamen Municipal Bureau of Science and Technology(3502Z20209005)Fundamental Research Funds for the Central Universities(20720200014,20720200069,20720190084)Program of Introducing Talents of Discipline to Universities(BP2018017)。
文摘Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),is characterized by a strong production of inflammatory cytokines such as TNF and IL-6,which underlie the severity of the disease.However,the molecular mechanisms responsible for such a strong immune response remains unclear.Here,utilizing targeted tandem mass spectrometry to analyze serum metabolome and lipidome in COVID-19 patients at different temporal stages,we identified that 611 metabolites(of 1,039)were significantly altered in COVID-19 patients.Among them,two metabolites,agmatine and putrescine,were prominently elevated in the serum of patients;and 2-quinolinecarboxylate was changed in a biphasic manner,elevated during early COVID-19 infection but levelled off.When tested in mouse embryonic fibroblasts(MEFs)and macrophages,these 3 metabolites were found to activate the NF-κB pathway that plays a pivotal role in governing cytokine production.Importantly,these metabolites were each able to cause strong increase of TNF and IL-6 levels when administered to wildtype mice,but not in the mice lacking NF-κB.Intriguingly,these metabolites have little effects on the activation of interferon regulatory factors(IRFs)for the production of type I interferons(IFNs)for antiviral defenses.These data suggest that circulating metabolites resulting from COVID-19 infection may act as effectors to elicit the peculiar systemic inflammatory responses,exhibiting severely strong proinflammatory cytokine production with limited induction of the interferons.Our study may provide a rationale for development of drugs to alleviate inflammation in COVID-19 patients.
基金NationalNatural Science Foundation of China,Grant/Award Numbers:81672440,31701156,81972625DICP,Grant/Award Number:ZZBS201803The Construction of Liaoning CancerResearch Center,Grant/Award Number:1564992449013。
文摘Background:Y-box binding protein 1(YB1 or YBX1)plays a critical role in tumorigenesis and cancer progression.However,whether YB1 affects malignant transformation by modulating non-codingRNAs remains largely unknown.This study aimed to investigate the relationship between YB1 and microRNAs and reveal the underlying mechanism by which YB1 impacts on tumor malignancy via miRNAs-mediated regulatory network.Methods:The biological functions of YB1 in hepatocellular carcinoma(HCC)cells were investigated by cell proliferation,wound healing,and transwell invasion assays.The miRNAs dysregulated by YB1 were screened by microarray analysis in HCC cell lines.The regulation of YB1 on miR-205 and miR-200b was determined by quantitative real-time PCR,dual-luciferase reporter assay,RNA immunoprecipitation,and pull-down assay.The relationships of YB1,DGCR8,Dicer,TUT4,and TUT1 were identified by pull-down and coimmunoprecipitation experiments.The cellular co-localization of YB1,DGCR8,and Dicer were detected by immunofluorescent staining.The in vivo effect of YB1 on tumor metastasis was determined by injecting MHCC97H cells transduced with YB1 shRNA or shControl via the tail vein in nude BALB/c mice.The expression levels of epithelial tomesenchymal transition markerswere detected by immunoblotting and immunohistochemistry assays.Results:YB1 promoted HCC cell migration and tumor metastasis by regulating miR-205/200b‒ZEB1 axis partially in a Snail-independent manner.YB1 suppressedmiR-205 and miR-200b maturation by interacting with the microprocessors DGCR8 and Dicer as well as TUT4 and TUT1 via the conserved cold shock domain.Subsequently,the downregulation of miR-205 and miR-200b enhanced ZEB1 expression,thus leading to increased cell migration and invasion.Furthermore,statistical analyses on gene expression data from HCC and normal liver tissues showed that YB1 expression was positively associated with ZEB1 expression and remarkably correlated with clinical prognosis.Conclusion:This study reveals a previously undescribed mechanism by which YB1 promotes cancer progression by regulating the miR-205/200b‒ZEB1 axis in HCC cells.Furthermore,these results highlight that YB1 may play biological functions via miRNAs-mediated gene regulation,and it can serve as a potential therapeutic target in human cancers.
基金supported by the National Key Research and Development Program of China(No.2022YFA0806503)the National Natural Science Foundation of China(No.81972625)+1 种基金the Dalian Science and Technology Innovation Funding(No.2019J12SN52)the Liaoning Revitalization Talents Program(No.XLYC2002035),China。
文摘Mammalian target of rapamycin(mTOR)controls cellular anabolism,and mTOR signaling is hyperactive in most cancer cells.As a result,inhibition of mTOR signaling benefits cancer patients.Rapamycin is a US Food and Drug Administration(FDA)-approved drug,a specific mTOR complex 1(mTORC1)inhibitor,for the treatment of several different types of cancer.However,rapamycin is reported to inhibit cancer growth rather than induce apoptosis.Pyruvate dehydrogenase complex(PDHc)is the gatekeeper for mitochondrial pyruvate oxidation.PDHc inactivation has been observed in a number of cancer cells,and this alteration protects cancer cells from senescence and nicotinamide adenine dinucleotide(NAD^(+))exhaustion.In this paper,we describe our finding that rapamycin treatment promotes pyruvate dehydrogenase E1 subunit alpha 1(PDHA1)phosphorylation and leads to PDHc inactivation dependent on mTOR signaling inhibition in cells.This inactivation reduces the sensitivity of cancer cells'response to rapamycin.As a result,rebooting PDHc activity with dichloroacetic acid(DCA),a pyruvate dehydrogenase kinase(PDK)inhibitor,promotes cancer cells'susceptibility to rapamycin treatment in vitro and in vivo.
基金the National Natural Science Foundation of China(Nos.81672440,21575142)Innovation Program of Science and Research from the DICP,CAS(No.DICP ZZBS201803)。
文摘Protein-metabolite interactions(PMIs)play important roles in various biological processes,especially in disease progression.However,due to the complexity of living cells,it is very difficult to identify specific PMIs.Herein,we chose one oncogenic factor,metadherin(MTDH),as a bait to identify its in vivo interacting metabolites in cancer cells.Cholesterol is an important metabolite and essential structural component of cell membranes.It could also drive several diseases including cancer.Interestingly,we found that cholesterol robustly interacted with MTDH and downregulated the expression of MTDH in cancer cells.Furthermore,MTDH disturbed metabolite alterations under cholesterol treatment in MTDH transduced cancer cells.Collectively,our results uncover an undescribed PMI where MTDH,as an oncogenic factor,might positively regulate cancer progression by interacting with choleste rol.This study interprets the theoretical basis of PMI-oriented cancer progression and targeting therapies in clinic.
基金This study was supported by the National Key Research and Development Program of China(2022YFA0806503)grants from the National Natural Science Foundation of China(No.81972625,No.21907093)+1 种基金Dalian Science and Technology Innovation Funding(2019J12SN52)Liaoning Revitalization Talents Program(XLYC2002035).
文摘Pyruvate is an essential fuel for maintaining the tricarboxylic acid(TCA)cycle in the mitochondria.However,the precise mole-cular mechanism of pyruvate uptake by mitochondrial pyruvate carrier(MPC)is largely unknown.Here,we report that the DNA/RNA-binding protein Y-box binding protein 1(YBX1)is localized to the mitochondrial inter-membrane space by its C-terminal domain(CTD)in cancer cells.In mitochondria,YBX1 inhibits pyruvate uptake by associating with MPC1/2,thereby suppressing pyruvate-dependent TCA cycle flux.This association,in turn,promotes MPC-mediated glutaminolysis and histone lactylation.Our findings reveal that the YBX1-MPC axis exhibits a positive correlation with metastatic potential,while does not affect cell proliferation in both cultured cells and tumor xenografts.Therefore,the restricted pyruvate uptake into mitochondria potentially represents a hallmark of metastatic capacity,suggesting that the YBX1-MPC axis is a therapeutic target for combating cancer metastasis.
基金This work was supported by the Scientific Instrument Developing Project of the Chinese Academy of Sciences(Grant No.YJKYYQ20190003)the Liao Ning Revitalization Talents Program(XLYC1802126)+1 种基金the Dalian City Foundation for Science and Technology Innovation(2019J12GX031)the National Natural Science Foundation of China(Grant Nos.21673237 and 21503224).
文摘Glutathione S-transferases(GSTs),detoxification enzymes that catalyze the addition of glutathione(GSH)to diverse electrophilic molecules,are often overexpressed in various tumor cells.While fluorescent probes for GSTs have often adopted the 2,4-dinitrobenzenesulfonyl(DNs)group as the receptor unit,they usually suffer from considerable background reaction noise with GSH due to excessive electron deficiency.However,weakening this reactivity is generally accompanied by loss of sensitivity for GSTs,and therefore,finely turning down the reactivity while maintaining certain sensitivity is critical for developing a practical probe.Here,we report a rational semiquantitative strategy for designing such a practical two-photon probe by introducing a parameter adopted from the conceptual density functional theory(CDFT),the local electrophilicityω_(k),to characterize this reactivity.As expected,kinetic studies establishedω_(k)as efficient to predict the reactivity with GSH,and probe NI3 showing the best performance was successfully applied to detecting GST activities in live cells and tissue sections with high sensitivity and signal-to-noise ratio.Photoinduced electron transfer of naphthalimide-based probes,captured by femtosecond transient absorption for the first time and unraveled by theoretical calculations,also contributes to the negligible background noise.
基金supported by the National Natural Science Foundation of China(No.81672440)Innovation Program of Science and Research from the DICP,CAS(No.DICP TMSR201601)the 100 Talents Program of Chinese Academy of Sciences
文摘Active endogenous metabolites regulate the viability of cells. This process is controlled by a series ofinteractions between small metabolites and large proteins. Previously, several studies had reported thatmetabolite regulates the protein functions, such as diacylglycerol to protein kinase C, lactose regulationof the lac repressor, and HIF-1α stabilization by 2-hydroxyglutarate. However, decades old traditionalbiochemical methods are insufficient to systematically investigate the bio-molecular reactions for a high-throughput discovery. Here, we have reviewed an update on the recently developed chemical proteomicscalled activity-based protein profiling （ABPP）. ABPP is able to identify proteins interacted eithercovalently or non-covalently with metabolites significantly. Thus, ABPP will facilitate the characteriza-tion of specific metabolite regulating; proteins in human disease progression.
基金supported by grants from the National Natural Science Foundation of China 82130071,82072516(Kanglai Tang)Sports Injury Repair and Reconstruction Research Innovative Team 41CZDH,Personalized Training Program of Leading Talent Training Object 414ZB2(Kanglai Tang)+4 种基金National Key R&D Program of China 2021YFC2401300(Lin Guo)National Natural Science Foundation of China 82173134(Hongming Miao)The Funding of Jinfeng Laboratory and Chongqing Outstanding Youth Fund CSTB2022NSCQ-JQX0010(Hongming Miao)NIH research grants R01AR062207,R01AR061484,R01AR076900,R01NS103931(Chuanju Liu)NIH research grants 1S10OD010582-01A1(Beatrix Ueberheide).
文摘Metabolites can double as a signaling modality that initiates physiological adaptations.Metabolism,a chemical language encoding biological information,has been recognized as a powerful principle directing inflammatory responses.Cytosolic pH is a regulator of inflammatory response in macrophages.Here,we found that L-malate exerts anti-inflammatory effect via BiP-IRF2BP2 signaling,which is a sensor of cytosolic pH in macrophages.First,L-malate,a TCA intermediate upregulated in pro-inflammatory macrophages,was identified as a potent anti-inflammatory metabolite through initial screening.Subsequent screening with DARTS and MS led to the isolation of L-malate-BiP binding.Further screening through protein‒protein interaction microarrays identified a L-malate-restrained coupling of BiP with IRF2BP2,a known anti-inflammatory protein.Interestingly,pH reduction,which promotes carboxyl protonation of L-malate,facilitates L-malate and carboxylate analogues such as succinate to bind BiP,and disrupt BiPIRF2BP2 interaction in a carboxyl-dependent manner.Both L-malate and acidification inhibit BiP-IRF2BP2 interaction,and protect IRF2BP2 from BiP-driven degradation in macrophages.Furthermore,both in vitro and in vivo,BiP-IRF2BP2 signal is required for effects of both L-malate and pH on inflammatory responses.Thesefindings reveal a previously unrecognized,proton/carboxylate dual sensing pathway wherein pH and L-malate regulate inflammatory responses,indicating the role of certain carboxylate metabolites as adaptors in the proton biosensing by interactions between macromolecules.
