Autophagy is an evolutionarily-conserved selfdegradative process that maintains cellular homeostasis by eliminating protein aggregates and damaged organelles.Recently, vesicle-associated membrane protein-associated pr...Autophagy is an evolutionarily-conserved selfdegradative process that maintains cellular homeostasis by eliminating protein aggregates and damaged organelles.Recently, vesicle-associated membrane protein-associated protein B(VAPB), which is associated with the familial form of amyotrophic lateral sclerosis, has been shown to regulate autophagy. In the present study, we demonstrated that knockdown of VAPB induced the up-regulation of beclin 1 expression, which promoted LC3(microtubuleassociated protein light chain 3) conversion and the formation of LC3 puncta, whereas overexpression of VAPB inhibited these processes. The regulation of beclin1 by VAPB was at the transcriptional level. Moreover,knockdown of VAPB increased autophagic flux, which promoted the degradation of the autophagy substrate p62 and neurodegenerative disease proteins. Our study provides evidence that the regulation of autophagy by VAPB is associated with the autophagy-initiating factor beclin 1.展开更多
Mono-ADP-ribosylation(MARylation)is a post-translational modification that regulates a variety of biological processes,including DNA damage repair,cell proliferation,metabolism,and stress and immune responses.In mamma...Mono-ADP-ribosylation(MARylation)is a post-translational modification that regulates a variety of biological processes,including DNA damage repair,cell proliferation,metabolism,and stress and immune responses.In mammals,MARylation is mainly catalyzed by ADP-ribosyltransferases(ARTs),which consist of two groups:ART cholera toxin-like(ARTCs)and ART diphtheria toxin-like(ARTDs,also known as PARPs).The human ARTC(hARTC)family is composed of four members:two active mono-ADP-ARTs(hARTC1 and hARTC5)and two enzymatically inactive enzymes(hARTC3 and hARTC4).In this study,we systematically examined the homology,expression,and localization pattern of the hARTC family,with a particular focus on hARTC1.Our results showed that hARTC3 interacted with hARTC1 and promoted the enzymatic activity of hARTC1 by stabilizing hARTC1.We also identified vesicle-associated membrane protein-associated protein B(VAPB)as a new target of hARTC1 and pinpointed Arg50 of VAPB as the ADP-ribosylation site.Furthermore,we demonstrated that knockdown of hARTC1 impaired intracellular calcium homeostasis,highlighting the functional importance of hARTC1-mediated VAPB Arg50 ADP-ribosylation in regulating calcium homeostasis.In summary,our study identified a new target of hARTC1 in the endoplasmic reticulum and suggested that ARTC1 plays a role in regulating calcium signaling.展开更多
为开发利用蕉柑落果资源,提高蕉柑落果中辛弗林的提取率,并尽量保护蕉柑组织形态完整,对质构保护液质量浓度以及真空气流细胞破壁技术的4个影响因素进行考察;并采用Box-Behnken试验设计及响应面分析对真空气流细胞破壁技术的工艺条件进...为开发利用蕉柑落果资源,提高蕉柑落果中辛弗林的提取率,并尽量保护蕉柑组织形态完整,对质构保护液质量浓度以及真空气流细胞破壁技术的4个影响因素进行考察;并采用Box-Behnken试验设计及响应面分析对真空气流细胞破壁技术的工艺条件进行优化。结果表明,质构保护最佳处理工艺为:首先在质量浓度0.3 g/100 m L的海藻酸钠溶液中浸泡20 min,接着在质量浓度0.15 g/100 m L的氯化钙溶液中浸泡60 min;真空气流细胞破壁前处理最佳工艺为:泄压温度100℃、压力差116 k Pa、停滞时间21 min、泄压3次。此时辛弗林提取率为69.42%,破果率为33.21%;相比直接进行亚临界水提取对照组,破果率下降了12.07%,辛弗林提取率增加了26.94%。展开更多
基金supported in part by the National Key R&D Program of China (2016YFC1306000)the National Natural Sciences Foundation of China (31330030, 31471012, and 81761148024)a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘Autophagy is an evolutionarily-conserved selfdegradative process that maintains cellular homeostasis by eliminating protein aggregates and damaged organelles.Recently, vesicle-associated membrane protein-associated protein B(VAPB), which is associated with the familial form of amyotrophic lateral sclerosis, has been shown to regulate autophagy. In the present study, we demonstrated that knockdown of VAPB induced the up-regulation of beclin 1 expression, which promoted LC3(microtubuleassociated protein light chain 3) conversion and the formation of LC3 puncta, whereas overexpression of VAPB inhibited these processes. The regulation of beclin1 by VAPB was at the transcriptional level. Moreover,knockdown of VAPB increased autophagic flux, which promoted the degradation of the autophagy substrate p62 and neurodegenerative disease proteins. Our study provides evidence that the regulation of autophagy by VAPB is associated with the autophagy-initiating factor beclin 1.
基金This work was supported by grants from the National Natural Science Foundation of China(32071277,82002594,81874160,and 32171295)the Natural Science Foundation of Hebei Province(C2021201012)+3 种基金S&T Program of Hebei(216Z2602G)the Interdisciplinary Research Program of Natural Science of Hebei University(DXK202006 and DXK202007)Hebei Natural Science Foundation for Outstanding Young Scholars(H2020201017)the High-level Talents Research Start-up Project of Hebei University(521000981352).
文摘Mono-ADP-ribosylation(MARylation)is a post-translational modification that regulates a variety of biological processes,including DNA damage repair,cell proliferation,metabolism,and stress and immune responses.In mammals,MARylation is mainly catalyzed by ADP-ribosyltransferases(ARTs),which consist of two groups:ART cholera toxin-like(ARTCs)and ART diphtheria toxin-like(ARTDs,also known as PARPs).The human ARTC(hARTC)family is composed of four members:two active mono-ADP-ARTs(hARTC1 and hARTC5)and two enzymatically inactive enzymes(hARTC3 and hARTC4).In this study,we systematically examined the homology,expression,and localization pattern of the hARTC family,with a particular focus on hARTC1.Our results showed that hARTC3 interacted with hARTC1 and promoted the enzymatic activity of hARTC1 by stabilizing hARTC1.We also identified vesicle-associated membrane protein-associated protein B(VAPB)as a new target of hARTC1 and pinpointed Arg50 of VAPB as the ADP-ribosylation site.Furthermore,we demonstrated that knockdown of hARTC1 impaired intracellular calcium homeostasis,highlighting the functional importance of hARTC1-mediated VAPB Arg50 ADP-ribosylation in regulating calcium homeostasis.In summary,our study identified a new target of hARTC1 in the endoplasmic reticulum and suggested that ARTC1 plays a role in regulating calcium signaling.
文摘为开发利用蕉柑落果资源,提高蕉柑落果中辛弗林的提取率,并尽量保护蕉柑组织形态完整,对质构保护液质量浓度以及真空气流细胞破壁技术的4个影响因素进行考察;并采用Box-Behnken试验设计及响应面分析对真空气流细胞破壁技术的工艺条件进行优化。结果表明,质构保护最佳处理工艺为:首先在质量浓度0.3 g/100 m L的海藻酸钠溶液中浸泡20 min,接着在质量浓度0.15 g/100 m L的氯化钙溶液中浸泡60 min;真空气流细胞破壁前处理最佳工艺为:泄压温度100℃、压力差116 k Pa、停滞时间21 min、泄压3次。此时辛弗林提取率为69.42%,破果率为33.21%;相比直接进行亚临界水提取对照组,破果率下降了12.07%,辛弗林提取率增加了26.94%。
