The E3 ubiquitin ligase,carboxyl terminus of heat shock protein 70(Hsp70)interacting protein(CHIP),also functions as a co-chaperone and plays a crucial role in the protein quality control system.In this study,we aimed...The E3 ubiquitin ligase,carboxyl terminus of heat shock protein 70(Hsp70)interacting protein(CHIP),also functions as a co-chaperone and plays a crucial role in the protein quality control system.In this study,we aimed to investigate the neuroprotective effect of overexpressed CHIP on Alzheimer’s disease.We used an adeno-associated virus vector that can cross the blood-brain barrier to mediate CHIP overexpression in APP/PS1 mouse brain.CHIP overexpression significantly ameliorated the performance of APP/PS1 mice in the Morris water maze and nest building tests,reduced amyloid-βplaques,and decreased the expression of both amyloid-βand phosphorylated tau.CHIP also alleviated the concentration of microglia and astrocytes around plaques.In APP/PS1 mice of a younger age,CHIP overexpression promoted an increase in ADAM10 expression and inhibitedβ-site APP cleaving enzyme 1,insulin degrading enzyme,and neprilysin expression.Levels of HSP70 and HSP40,which have functional relevance to CHIP,were also increased.Single nuclei transcriptome sequencing in the hippocampus of CHIP overexpressed mice showed that the lysosomal pathway and oligodendrocyte-related biological processes were up-regulated,which may also reflect a potential mechanism for the neuroprotective effect of CHIP.Our research shows that CHIP effectively reduces the behavior and pathological manifestations of APP/PS1 mice.Indeed,overexpression of CHIP could be a beneficial approach for the treatment of Alzheimer’s disease.展开更多
热激蛋白HSP70(heat shock protein 70)家族是一类结构高度保守和功能强大的蛋白家族,然而有关其在二倍体长穗偃麦草(Thinopyrum elongatum)中的研究尚未报道。为研究二倍体长穗偃麦草HSP70基因家族(TeHSP70),基于生物信息学方法对二倍...热激蛋白HSP70(heat shock protein 70)家族是一类结构高度保守和功能强大的蛋白家族,然而有关其在二倍体长穗偃麦草(Thinopyrum elongatum)中的研究尚未报道。为研究二倍体长穗偃麦草HSP70基因家族(TeHSP70),基于生物信息学方法对二倍体长穗偃麦草TeHSP70基因进行鉴定,并系统分析其在盐胁迫和多组织中的表达模式。结果表明,在二倍体长穗偃麦草中共鉴定到29个TeHSP70基因,可分为3个亚族,均匀地分布在7条染色体上;其中有26个TeHSP70基因定位于细胞质,且都包含α-螺旋、延伸链、β-折叠和无规则卷曲结构。29个TeHSP70基因在基序上高度保守,但基因结构存在差异,此外在各基因上游均包含多种激素和胁迫响应的顺式作用元件。共线性分析显示,29个TeHSP70基因间存在3对串联重复和片段重复事件;同时与水稻和小麦有着多对同源基因,而与拟南芥同源基因较少。蛋白质互作网络分析显示,有11个TeHSP70蛋白间存在复杂且密切的蛋白质互作网络。表达分析显示,随着盐胁迫的增加(0~300 mmol·L^(-1))有7个TeHSP70基因持续上调,有6个TeHSP70基因持续下调,其余16个TeHSP70基因未表达或变化特征不连续;有25个TeHSP70基因在10个不同组织中均有表达,有4个TeHSP70s基因仅在部分组织中表达。以上研究结果为深入解析二倍体长穗偃麦草HSP70功能提供了有效的借鉴和参考。展开更多
为探究中华卤虫(Artemia sinica)热激蛋白HSP70的分子功能及其在生态适应中的作用机制,文章以运城盐湖中华卤虫为研究对象,通过RT-PCR扩增获得AsHSP70基因完整开放阅读框(ORF)。将目的片段克隆至pGEX-4T-1表达载体,转化BL21(DE3)plySs...为探究中华卤虫(Artemia sinica)热激蛋白HSP70的分子功能及其在生态适应中的作用机制,文章以运城盐湖中华卤虫为研究对象,通过RT-PCR扩增获得AsHSP70基因完整开放阅读框(ORF)。将目的片段克隆至pGEX-4T-1表达载体,转化BL21(DE3)plySs感受态细胞。SDS-PAGE分析显示,IPTG诱导后可成功表达约98.3 kDa的GST-AsHSP70融合蛋白。通过优化表达条件,发现37℃、1 mM IPTG诱导4 h总蛋白表达量最高;25℃、0.5 mM IPTG诱导4 h可显著提高蛋白产量。文章建立了运城盐湖卤虫AsHSP70基因的高效原核表达体系,为后续解析HSP70在极端环境适应中的分子机制以及开发其作为免疫佐剂的潜在应用提供了技术基础。展开更多
基金supported by the National Natural Science Foundation of China,Nos.91849115 and U1904207(to YX),81974211 and 82171247(to CS)Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences,No.2020-PT310-01(to YX).
文摘The E3 ubiquitin ligase,carboxyl terminus of heat shock protein 70(Hsp70)interacting protein(CHIP),also functions as a co-chaperone and plays a crucial role in the protein quality control system.In this study,we aimed to investigate the neuroprotective effect of overexpressed CHIP on Alzheimer’s disease.We used an adeno-associated virus vector that can cross the blood-brain barrier to mediate CHIP overexpression in APP/PS1 mouse brain.CHIP overexpression significantly ameliorated the performance of APP/PS1 mice in the Morris water maze and nest building tests,reduced amyloid-βplaques,and decreased the expression of both amyloid-βand phosphorylated tau.CHIP also alleviated the concentration of microglia and astrocytes around plaques.In APP/PS1 mice of a younger age,CHIP overexpression promoted an increase in ADAM10 expression and inhibitedβ-site APP cleaving enzyme 1,insulin degrading enzyme,and neprilysin expression.Levels of HSP70 and HSP40,which have functional relevance to CHIP,were also increased.Single nuclei transcriptome sequencing in the hippocampus of CHIP overexpressed mice showed that the lysosomal pathway and oligodendrocyte-related biological processes were up-regulated,which may also reflect a potential mechanism for the neuroprotective effect of CHIP.Our research shows that CHIP effectively reduces the behavior and pathological manifestations of APP/PS1 mice.Indeed,overexpression of CHIP could be a beneficial approach for the treatment of Alzheimer’s disease.
文摘为探究中华卤虫(Artemia sinica)热激蛋白HSP70的分子功能及其在生态适应中的作用机制,文章以运城盐湖中华卤虫为研究对象,通过RT-PCR扩增获得AsHSP70基因完整开放阅读框(ORF)。将目的片段克隆至pGEX-4T-1表达载体,转化BL21(DE3)plySs感受态细胞。SDS-PAGE分析显示,IPTG诱导后可成功表达约98.3 kDa的GST-AsHSP70融合蛋白。通过优化表达条件,发现37℃、1 mM IPTG诱导4 h总蛋白表达量最高;25℃、0.5 mM IPTG诱导4 h可显著提高蛋白产量。文章建立了运城盐湖卤虫AsHSP70基因的高效原核表达体系,为后续解析HSP70在极端环境适应中的分子机制以及开发其作为免疫佐剂的潜在应用提供了技术基础。
