Non-specific lipid transfer proteins(nsLTPs) are small, basic proteins that are characterized by an eight-cysteine motif. The biological functions of these proteins have been reported to involve plant reproduction and...Non-specific lipid transfer proteins(nsLTPs) are small, basic proteins that are characterized by an eight-cysteine motif. The biological functions of these proteins have been reported to involve plant reproduction and biotic or abiotic stress response. With the completion of the barley genome sequence, a genome-wide analysis of nsLTPs in barley(Hordeum vulgare L.)(HvLTPs) will be helpful for understanding the function of nsLTPs in plants. We performed a genome-wide analysis of the nsLTP gene family in barley and identified 70 nsLTP genes,which can be divided into five types(1, 2, C, D, and G). Each type of nsLTPs shares similar exon and intron gene structures. Expression analysis showed that barley nsLTPs have diverse expression patterns, revealing their various roles. Our results shed light on the phylogenetic relationships and potential functions of barley nsLTPs and will be useful for future studies of barley development and molecular breeding.展开更多
Plant nonspecific lipid transfer proteins (nsLTPs) are widely distributed through plant kingdom and are characterized by the presence of a central hydrophobic cavity, suitable for binding various hydrophobic molecules...Plant nonspecific lipid transfer proteins (nsLTPs) are widely distributed through plant kingdom and are characterized by the presence of a central hydrophobic cavity, suitable for binding various hydrophobic molecules. Despite extensive research on nsLTP in different plant species, mostly angiosperm, and the great diversity of physiological processes in which they seem to be involved, their exact functions still remain unclear. Also, very limited experimental data are available on nsLTP in gymnosperm. In this study, we report for the first time on the molecular cloning of nsLTP, from Pinus sylvestris L.(PsLTP1, GenBank accession JN980402.1) and the expression pattern of PsLTP1 during ontogenesis and in response to environmental stress conditions. Total RNA from roots of 7-day old pine seedlings was used to isolate the cDNA clone, corresponding to Scots pine lipid transfer protein. The open reading frame of PsLTP1 consists of 372 bp encoding a protein of 123 amino acids. Amino acid sequence alignment revealed that mature PsLTP1 shares high level of similarity with nsLTP from other conifers and with well-studied nsLTPs from angiosperms. The PsLTP1 contains a 27-amino-acid N-terminal signal sequence and presents all the features of a plant nsLTP. Amino acid comparison analysis and 3D structure prediction showed that PsLTP1 is a type 1 nsLTP. The results of the expression analysis of Scots pine PsLTP1 gene revealed that its transcripts accumulate in actively growing tissues. Furthermore, transcription of PsLTP1 was upregulated in response to cold and salt treatments, and downregulated during acidic, osmotic and water stresses.展开更多
A plant＇s capability to cope with environmental challenges largely relies on signal transmission through mitogen-activated protein kinase （MAPK） cascades. In Arabidopsis thaliana, MPK3 is particularly strongly asso...A plant＇s capability to cope with environmental challenges largely relies on signal transmission through mitogen-activated protein kinase （MAPK） cascades. In Arabidopsis thaliana, MPK3 is particularly strongly associated with numerous abiotic and biotic stress responses. Identification of MPK3 substrates is a milestone towards improving stress resistance in plants. Here, we characterize AZI1, a lipid transfer protein （LTP）-related hybrid proline-rich protein （HyPRP）, as a novel target of MPK3. AZI1 is phosphorylated by MPK3 in vitro. As documented by co-immunoprecipitation and bimolecular fluorescence complementation experiments, AZI1 interacts with MPK3 to form protein complexes in planta. Furthermore, null mutants of azil are hypersensitive to salt stress, while AZIl-overexpressing lines are markedly more tolerant. AZI1 overexpression in the mpk3 genetic background partially alleviates the salt-hypersensitive phenotype of this mutant, but functional MPK3 appears to be required for the full extent of AZIl-conferred robustness. Notably, this robustness does not come at the expense of normal development. Immunoblot and RT-PCR data point to a role of MPK3 as positive regulator of AZI1 abundance.展开更多
Objective： Plasma phospholipid transfer protein （PLTP） is a key determinant of lipoprotein metabolism, and both animal and human studies converge to indicate that PLTP promotes atherogenesis and its thromboembolic ...Objective： Plasma phospholipid transfer protein （PLTP） is a key determinant of lipoprotein metabolism, and both animal and human studies converge to indicate that PLTP promotes atherogenesis and its thromboembolic complications. Moreover, it has recently been reported that PLTP modulates inflammation and immune responses. Although earlier studies from our group demonstrated that PLTP can modify macrophage activation, the implication of PLTP in the modulation of T-cell-mediated immune responses has never been investigated and was therefore addressed in the present study. Approach and results： In the present study, we demonstrated that PLTP deficiency in mice has a profound effect on CD4＋ ThO cell polarization, with a shift towards the anti-inflammatory Th2 phenotype under both normal and pathological conditions. In a model of contact hypersensitivity, a significantly impaired response to skin sensitization with the hapten-2,4-dinitrofluorobenzene （DNFB） was observed in PLTP-deficient mice compared to wild-type （WT） mice. Interestingly, PLTP deficiency in mice exerted no effect on the counts of total white blood cells, lymphocytes, granulocytes, or monocytes in the peripheral blood. Moreover, PLTP deficiency did not modify the amounts of CD4＋ and CD8＋ T lymphocyte subsets. However, PLTP-deficiency, associated with upregulation of the Th2 phenotype, was accompanied by a significant decrease in the production of the pro-Thl cytokine interleukin 18 by accessory cells. Conclusions： For the first time, this work reports a physiological role for PLTP in the polarization of CD4＋ T cells toward the pro-inflammatory Th I phenotype.展开更多
In plants,lipid transfer proteins(LTPs)transport pollen wall constituents from the tapetum to the exine,a process essential for pollen wall development.However,the functional cooperation of different LTPs in pollen wa...In plants,lipid transfer proteins(LTPs)transport pollen wall constituents from the tapetum to the exine,a process essential for pollen wall development.However,the functional cooperation of different LTPs in pollen wall development is not well understood.In this study,we have identified and characterized a grassspecific LTP gene,Os LTP47,an important regulator of pollen wall formation in rice(Oryza sativa).Os LTP47 encodes a membrane-localized LTP and in vitro lipid-binding assays confirms that Os LTP47 has lipidbinding activity.Dysfunction of Os LTP47 causes disordered lipid metabolism and defective pollen walls,leading to male sterility.Yeast two-hybrid and pull-down assays reveal that Os LTP47 physically interacts with another LTP,Os C6.These findings suggest that the plasma membrane-localized Os LTP47 may function as a mediator in a lipid transfer relay through association with cytosolic and/or locular Os C6 for pollen wall development and that various LTPs may function in a coordinated manner to transport lipid molecules during pollen wall development.展开更多
非特异性脂质转移蛋白(nsLTP,non-specific lipid transfer proteins)在植物脂质转运和分泌中发挥重要作用。本研究从薰衣草(Lavandula angustifolia)中克隆到2个II型nsLTP基因,命名为nsLTP2-1和nsLTP2-2,并对其进行功能分析。生信分析...非特异性脂质转移蛋白(nsLTP,non-specific lipid transfer proteins)在植物脂质转运和分泌中发挥重要作用。本研究从薰衣草(Lavandula angustifolia)中克隆到2个II型nsLTP基因,命名为nsLTP2-1和nsLTP2-2,并对其进行功能分析。生信分析表明,nsLTP2-1和ns LTP2-2分别编码119个和117个氨基酸,具有脂转移蛋白(LTP,lipid transfer proteins)保守结构域和8个高度保守的半胱氨酸残基;系统进化分析显示它们处于两个分支,与同科的紫苏(Perilla frutescens)相似性最高。基因表达分析显示2个基因均在花蕾中高表达,在叶片、茎和花瓣中几乎不表达,在花萼中的表达存在差异,nsLTP2-1和nsLTP2-2分别在成熟花萼和幼嫩花萼中表达量更高;2个基因在花蕾和叶片中的表达均受到强光诱导,且在花蕾中的表达均受脱落酸诱导,而叶片中nsLTP2-1和nsLTP2-2的表达分别受茉莉酸甲酯和乙烯诱导。亚细胞定位显示2个nsLTPs均定位在细胞膜和细胞壁上,可能与次生代谢物的转运有关。过表达nsLTP2-1和nsLTP2-2烟草叶片经尼罗红染色后,经485~543 nm激发光激发,叶片腺毛头部的荧光显示多于野生型,说明本研究中的nsLTPs可能在脂类的合成和转运中起重要作用。这些结果为明确薰衣草脂转移蛋白在脂类及萜类转运中的功能研究提供了参考。展开更多
磷脂转运蛋白(phospholipid transfer protein, PLTP)是循环内介导脂蛋白代谢的重要蛋白之一,其血浆活性与多种代谢性疾病或动脉粥样硬化性心血管疾病相关。本文就PLTP的生物学特性及其与糖脂代谢、动脉粥样硬化和血栓等病理过程相关的...磷脂转运蛋白(phospholipid transfer protein, PLTP)是循环内介导脂蛋白代谢的重要蛋白之一,其血浆活性与多种代谢性疾病或动脉粥样硬化性心血管疾病相关。本文就PLTP的生物学特性及其与糖脂代谢、动脉粥样硬化和血栓等病理过程相关的研究进展、研究前景和临床潜在应用进行介绍。展开更多
利用hiTAIL-PCR法得到了2个蜡梅(Chimonanthus praecox)非特异性脂转移蛋白(non-specific lipid transfer protein)基因家族成员CpLTP3和CpLTP4翻译起始位点上游启动子序列,长度分别为1298bp和838bp。生物信息学分析表明2个序列均存在...利用hiTAIL-PCR法得到了2个蜡梅(Chimonanthus praecox)非特异性脂转移蛋白(non-specific lipid transfer protein)基因家族成员CpLTP3和CpLTP4翻译起始位点上游启动子序列,长度分别为1298bp和838bp。生物信息学分析表明2个序列均存在启动子的基本元件TATA-box和CAAT-box及多个与植物非生物胁迫相关的响应元件。在烟草叶片中的瞬时表达结果表明这两个启动子序列均具备驱动报告基因GUS表达的功能。展开更多
利用RACE技术从板栗短雄花序突变体中扩增得到660bp的板栗脂质转运蛋白(lipid transfer protein,LTP)cDNA片段。该cDNA编码118个氨基酸,具有8个位置保守的半胱氨酸(C)残基及26个氨基酸的信号肽,它与棉花、草莓脂质转运蛋白氨基酸序列相...利用RACE技术从板栗短雄花序突变体中扩增得到660bp的板栗脂质转运蛋白(lipid transfer protein,LTP)cDNA片段。该cDNA编码118个氨基酸,具有8个位置保守的半胱氨酸(C)残基及26个氨基酸的信号肽,它与棉花、草莓脂质转运蛋白氨基酸序列相似性为63%,基因序列已提交数据库(GenBank),其登录号分别为FJ490676(基因)和ACL01093(蛋白)。荧光定量分析表明板栗短雄花序突变体较正常雄花序的脂质转运蛋白表达量更高。将板栗脂质转运蛋白基因插入到硫氧还蛋白融合表达载体pET32a(+)中,构建了板栗的原核表达载体pET-LTP,在大肠杆菌菌株Rosetta-gamiTM2(DE3)中,IPTG诱导5h大量表达约为30ku的融合蛋白,通过Ni2+-chelating sepharose fast flow柱纯化的融合蛋白具有抑制板栗镰刀菌属病原真菌孢子萌发的功能。展开更多
基金supported by funds from the National Key Research and Development Program of China (2016YFD0100903)
文摘Non-specific lipid transfer proteins(nsLTPs) are small, basic proteins that are characterized by an eight-cysteine motif. The biological functions of these proteins have been reported to involve plant reproduction and biotic or abiotic stress response. With the completion of the barley genome sequence, a genome-wide analysis of nsLTPs in barley(Hordeum vulgare L.)(HvLTPs) will be helpful for understanding the function of nsLTPs in plants. We performed a genome-wide analysis of the nsLTP gene family in barley and identified 70 nsLTP genes,which can be divided into five types(1, 2, C, D, and G). Each type of nsLTPs shares similar exon and intron gene structures. Expression analysis showed that barley nsLTPs have diverse expression patterns, revealing their various roles. Our results shed light on the phylogenetic relationships and potential functions of barley nsLTPs and will be useful for future studies of barley development and molecular breeding.
基金supported by grants from the Ministry of Education and Science of Ukraine(0116U003593)grant from cieA3(Campus de Excelencia Internacional Agroalimentario)-UCO,Spain
文摘Plant nonspecific lipid transfer proteins (nsLTPs) are widely distributed through plant kingdom and are characterized by the presence of a central hydrophobic cavity, suitable for binding various hydrophobic molecules. Despite extensive research on nsLTP in different plant species, mostly angiosperm, and the great diversity of physiological processes in which they seem to be involved, their exact functions still remain unclear. Also, very limited experimental data are available on nsLTP in gymnosperm. In this study, we report for the first time on the molecular cloning of nsLTP, from Pinus sylvestris L.(PsLTP1, GenBank accession JN980402.1) and the expression pattern of PsLTP1 during ontogenesis and in response to environmental stress conditions. Total RNA from roots of 7-day old pine seedlings was used to isolate the cDNA clone, corresponding to Scots pine lipid transfer protein. The open reading frame of PsLTP1 consists of 372 bp encoding a protein of 123 amino acids. Amino acid sequence alignment revealed that mature PsLTP1 shares high level of similarity with nsLTP from other conifers and with well-studied nsLTPs from angiosperms. The PsLTP1 contains a 27-amino-acid N-terminal signal sequence and presents all the features of a plant nsLTP. Amino acid comparison analysis and 3D structure prediction showed that PsLTP1 is a type 1 nsLTP. The results of the expression analysis of Scots pine PsLTP1 gene revealed that its transcripts accumulate in actively growing tissues. Furthermore, transcription of PsLTP1 was upregulated in response to cold and salt treatments, and downregulated during acidic, osmotic and water stresses.
文摘A plant＇s capability to cope with environmental challenges largely relies on signal transmission through mitogen-activated protein kinase （MAPK） cascades. In Arabidopsis thaliana, MPK3 is particularly strongly associated with numerous abiotic and biotic stress responses. Identification of MPK3 substrates is a milestone towards improving stress resistance in plants. Here, we characterize AZI1, a lipid transfer protein （LTP）-related hybrid proline-rich protein （HyPRP）, as a novel target of MPK3. AZI1 is phosphorylated by MPK3 in vitro. As documented by co-immunoprecipitation and bimolecular fluorescence complementation experiments, AZI1 interacts with MPK3 to form protein complexes in planta. Furthermore, null mutants of azil are hypersensitive to salt stress, while AZIl-overexpressing lines are markedly more tolerant. AZI1 overexpression in the mpk3 genetic background partially alleviates the salt-hypersensitive phenotype of this mutant, but functional MPK3 appears to be required for the full extent of AZIl-conferred robustness. Notably, this robustness does not come at the expense of normal development. Immunoblot and RT-PCR data point to a role of MPK3 as positive regulator of AZI1 abundance.
文摘Objective： Plasma phospholipid transfer protein （PLTP） is a key determinant of lipoprotein metabolism, and both animal and human studies converge to indicate that PLTP promotes atherogenesis and its thromboembolic complications. Moreover, it has recently been reported that PLTP modulates inflammation and immune responses. Although earlier studies from our group demonstrated that PLTP can modify macrophage activation, the implication of PLTP in the modulation of T-cell-mediated immune responses has never been investigated and was therefore addressed in the present study. Approach and results： In the present study, we demonstrated that PLTP deficiency in mice has a profound effect on CD4＋ ThO cell polarization, with a shift towards the anti-inflammatory Th2 phenotype under both normal and pathological conditions. In a model of contact hypersensitivity, a significantly impaired response to skin sensitization with the hapten-2,4-dinitrofluorobenzene （DNFB） was observed in PLTP-deficient mice compared to wild-type （WT） mice. Interestingly, PLTP deficiency in mice exerted no effect on the counts of total white blood cells, lymphocytes, granulocytes, or monocytes in the peripheral blood. Moreover, PLTP deficiency did not modify the amounts of CD4＋ and CD8＋ T lymphocyte subsets. However, PLTP-deficiency, associated with upregulation of the Th2 phenotype, was accompanied by a significant decrease in the production of the pro-Thl cytokine interleukin 18 by accessory cells. Conclusions： For the first time, this work reports a physiological role for PLTP in the polarization of CD4＋ T cells toward the pro-inflammatory Th I phenotype.
基金supported by grants from the Guangdong Natural Science Funds for Distinguished Young Scholars (2021B1515020089)the National Natural Science Foundation of China (32030080)the Major Program of Guangdong Basic and Applied Research(2019B030302006)
文摘In plants,lipid transfer proteins(LTPs)transport pollen wall constituents from the tapetum to the exine,a process essential for pollen wall development.However,the functional cooperation of different LTPs in pollen wall development is not well understood.In this study,we have identified and characterized a grassspecific LTP gene,Os LTP47,an important regulator of pollen wall formation in rice(Oryza sativa).Os LTP47 encodes a membrane-localized LTP and in vitro lipid-binding assays confirms that Os LTP47 has lipidbinding activity.Dysfunction of Os LTP47 causes disordered lipid metabolism and defective pollen walls,leading to male sterility.Yeast two-hybrid and pull-down assays reveal that Os LTP47 physically interacts with another LTP,Os C6.These findings suggest that the plasma membrane-localized Os LTP47 may function as a mediator in a lipid transfer relay through association with cytosolic and/or locular Os C6 for pollen wall development and that various LTPs may function in a coordinated manner to transport lipid molecules during pollen wall development.
文摘非特异性脂质转移蛋白(nsLTP,non-specific lipid transfer proteins)在植物脂质转运和分泌中发挥重要作用。本研究从薰衣草(Lavandula angustifolia)中克隆到2个II型nsLTP基因,命名为nsLTP2-1和nsLTP2-2,并对其进行功能分析。生信分析表明,nsLTP2-1和ns LTP2-2分别编码119个和117个氨基酸,具有脂转移蛋白(LTP,lipid transfer proteins)保守结构域和8个高度保守的半胱氨酸残基;系统进化分析显示它们处于两个分支,与同科的紫苏(Perilla frutescens)相似性最高。基因表达分析显示2个基因均在花蕾中高表达,在叶片、茎和花瓣中几乎不表达,在花萼中的表达存在差异,nsLTP2-1和nsLTP2-2分别在成熟花萼和幼嫩花萼中表达量更高;2个基因在花蕾和叶片中的表达均受到强光诱导,且在花蕾中的表达均受脱落酸诱导,而叶片中nsLTP2-1和nsLTP2-2的表达分别受茉莉酸甲酯和乙烯诱导。亚细胞定位显示2个nsLTPs均定位在细胞膜和细胞壁上,可能与次生代谢物的转运有关。过表达nsLTP2-1和nsLTP2-2烟草叶片经尼罗红染色后,经485~543 nm激发光激发,叶片腺毛头部的荧光显示多于野生型,说明本研究中的nsLTPs可能在脂类的合成和转运中起重要作用。这些结果为明确薰衣草脂转移蛋白在脂类及萜类转运中的功能研究提供了参考。
文摘磷脂转运蛋白(phospholipid transfer protein, PLTP)是循环内介导脂蛋白代谢的重要蛋白之一,其血浆活性与多种代谢性疾病或动脉粥样硬化性心血管疾病相关。本文就PLTP的生物学特性及其与糖脂代谢、动脉粥样硬化和血栓等病理过程相关的研究进展、研究前景和临床潜在应用进行介绍。
文摘利用hiTAIL-PCR法得到了2个蜡梅(Chimonanthus praecox)非特异性脂转移蛋白(non-specific lipid transfer protein)基因家族成员CpLTP3和CpLTP4翻译起始位点上游启动子序列,长度分别为1298bp和838bp。生物信息学分析表明2个序列均存在启动子的基本元件TATA-box和CAAT-box及多个与植物非生物胁迫相关的响应元件。在烟草叶片中的瞬时表达结果表明这两个启动子序列均具备驱动报告基因GUS表达的功能。
文摘利用RACE技术从板栗短雄花序突变体中扩增得到660bp的板栗脂质转运蛋白(lipid transfer protein,LTP)cDNA片段。该cDNA编码118个氨基酸,具有8个位置保守的半胱氨酸(C)残基及26个氨基酸的信号肽,它与棉花、草莓脂质转运蛋白氨基酸序列相似性为63%,基因序列已提交数据库(GenBank),其登录号分别为FJ490676(基因)和ACL01093(蛋白)。荧光定量分析表明板栗短雄花序突变体较正常雄花序的脂质转运蛋白表达量更高。将板栗脂质转运蛋白基因插入到硫氧还蛋白融合表达载体pET32a(+)中,构建了板栗的原核表达载体pET-LTP,在大肠杆菌菌株Rosetta-gamiTM2(DE3)中,IPTG诱导5h大量表达约为30ku的融合蛋白,通过Ni2+-chelating sepharose fast flow柱纯化的融合蛋白具有抑制板栗镰刀菌属病原真菌孢子萌发的功能。
