Ultraviolet radiation by its wavelength is divided into: UVA, UVB and UVC. Only UVA and UVB manage to penetrate the ozone layer, but due to anthropological activities, all of them are capable of interacting with human...Ultraviolet radiation by its wavelength is divided into: UVA, UVB and UVC. Only UVA and UVB manage to penetrate the ozone layer, but due to anthropological activities, all of them are capable of interacting with humans to a greater or lesser extent, and can generate adverse effects such as cellular stress when interacting with intra-and extracellular biomolecules. The skin is the first organ in contact with UV radiation, and the stress it generates can be analyzed by the expression of a bioindicator of cellular damage such as Hsp70. Therefore, the objective of the project was: to determine the effect of UVA, UVB and UVC radiation on HaCaT epithelial cells, by analyzing the expression of Hsp70. Materials and methods: HaCaT cells were cultured in vitro, which were irradiated with UVA, UVB and UVC light at different doses, to subsequently determine the degree of Hsp70 expression by Immunodetection by PAGE-SDS and Western Blot. Results: Basal expression of Hsp70 was observed in no irradiated HaCaT cells. When HaCaT cells were irradiated with UVA, UVB, UVC, an increase in this Hsp70 protein was observed. With UVA, a higher degree of expression was observed at a time of 30 minutes of irradiation. With UVB the highest expression shifted to a time of 20 minutes. With UVC, overexpression was observed after 10 minutes. Conclusion: UV radiation generates cellular stress on HaCaT cells, evaluated by the stress bioindicator Hsp70. According to the wavelength of UV radiation, those that have a shorter wavelength have a greater potential for cellular damage, such as UVC.展开更多
为了研究黄鳝(Monopterus albus)热休克蛋白70(Heat Shock Protein 70,简称HSP70)的结构和功能,试验采用生物信息学方法对黄鳝HSP70的基本理化性质、亲/疏水性、信号肽、磷酸化位点、糖基化、无序化特征、二级结构、三级结构、相互作用...为了研究黄鳝(Monopterus albus)热休克蛋白70(Heat Shock Protein 70,简称HSP70)的结构和功能,试验采用生物信息学方法对黄鳝HSP70的基本理化性质、亲/疏水性、信号肽、磷酸化位点、糖基化、无序化特征、二级结构、三级结构、相互作用蛋白网络及进化树进行了研究。结果表明,黄鳝HSP70由441个氨基酸组成,相对分子质量为48146.31,等电点为5.98,整体呈碱性,是较稳定的亲水性蛋白质,有信号肽,但不存在跨膜结构,属于分泌型蛋白质。黄鳝HSP70存在35个潜在磷酸化位点,其中包括19个Ser、14个Thr和2个Tyr磷酸化位点,有5个典型的N糖基化位点。二级结构以α-螺旋(39.68%)、延伸链(20.41%)、β-折叠(6.58%)和无规则卷曲(33.33%)为主要结构。热休克蛋白70(HSP70)作为分子伴侣,在蛋白质折叠和运输、细胞周期调控、细胞凋亡和精子发生等方面发挥着重要作用。研究表明,黄鳝HSP70在抵抗外界应激源、环境应激保护等方面起着重要作用。展开更多
Protein folding in crowding cellular environment often relies on the assistance of various chaperones. Hsp70 is one of the most ubiquitous chaperones in cells. Previous studies showed that the chaperone–client intera...Protein folding in crowding cellular environment often relies on the assistance of various chaperones. Hsp70 is one of the most ubiquitous chaperones in cells. Previous studies showed that the chaperone–client interactions at the open state tend to remodel the protein folding energy landscape and direct the protein folding as a foldase. In this work, we further investigate how the chaperone–client interaction strength modulates the foldase function of Hsp70 by using molecular simulations. The results showed that the time of substrate folding(including the whole folding step and substrate release step) has a non-monotonic dependence on the interaction strength. With the increasing of the chaperone–client interaction strength, the folding time decreases first, and then increases. More detailed analysis showed that when the chaperone–client interaction is too strong, even small number of chaperones–client contacts can maintain the substrate bound with the chaperone. The sampling of the transient chaperones–client complex with sparse inter-molecule contacts makes the client protein have chance to access the misfolded state even it is bound with chaperone. The current results suggest that the interaction strength is an important factor controlling the Hsp70 chaperoning function.展开更多
文摘Ultraviolet radiation by its wavelength is divided into: UVA, UVB and UVC. Only UVA and UVB manage to penetrate the ozone layer, but due to anthropological activities, all of them are capable of interacting with humans to a greater or lesser extent, and can generate adverse effects such as cellular stress when interacting with intra-and extracellular biomolecules. The skin is the first organ in contact with UV radiation, and the stress it generates can be analyzed by the expression of a bioindicator of cellular damage such as Hsp70. Therefore, the objective of the project was: to determine the effect of UVA, UVB and UVC radiation on HaCaT epithelial cells, by analyzing the expression of Hsp70. Materials and methods: HaCaT cells were cultured in vitro, which were irradiated with UVA, UVB and UVC light at different doses, to subsequently determine the degree of Hsp70 expression by Immunodetection by PAGE-SDS and Western Blot. Results: Basal expression of Hsp70 was observed in no irradiated HaCaT cells. When HaCaT cells were irradiated with UVA, UVB, UVC, an increase in this Hsp70 protein was observed. With UVA, a higher degree of expression was observed at a time of 30 minutes of irradiation. With UVB the highest expression shifted to a time of 20 minutes. With UVC, overexpression was observed after 10 minutes. Conclusion: UV radiation generates cellular stress on HaCaT cells, evaluated by the stress bioindicator Hsp70. According to the wavelength of UV radiation, those that have a shorter wavelength have a greater potential for cellular damage, such as UVC.
文摘为了研究黄鳝(Monopterus albus)热休克蛋白70(Heat Shock Protein 70,简称HSP70)的结构和功能,试验采用生物信息学方法对黄鳝HSP70的基本理化性质、亲/疏水性、信号肽、磷酸化位点、糖基化、无序化特征、二级结构、三级结构、相互作用蛋白网络及进化树进行了研究。结果表明,黄鳝HSP70由441个氨基酸组成,相对分子质量为48146.31,等电点为5.98,整体呈碱性,是较稳定的亲水性蛋白质,有信号肽,但不存在跨膜结构,属于分泌型蛋白质。黄鳝HSP70存在35个潜在磷酸化位点,其中包括19个Ser、14个Thr和2个Tyr磷酸化位点,有5个典型的N糖基化位点。二级结构以α-螺旋(39.68%)、延伸链(20.41%)、β-折叠(6.58%)和无规则卷曲(33.33%)为主要结构。热休克蛋白70(HSP70)作为分子伴侣,在蛋白质折叠和运输、细胞周期调控、细胞凋亡和精子发生等方面发挥着重要作用。研究表明,黄鳝HSP70在抵抗外界应激源、环境应激保护等方面起着重要作用。
基金Project supported by the National Natural Science Foundation of China (Grant Nos. 11305139 and 11974173)the HPC Center of Nanjing University。
文摘Protein folding in crowding cellular environment often relies on the assistance of various chaperones. Hsp70 is one of the most ubiquitous chaperones in cells. Previous studies showed that the chaperone–client interactions at the open state tend to remodel the protein folding energy landscape and direct the protein folding as a foldase. In this work, we further investigate how the chaperone–client interaction strength modulates the foldase function of Hsp70 by using molecular simulations. The results showed that the time of substrate folding(including the whole folding step and substrate release step) has a non-monotonic dependence on the interaction strength. With the increasing of the chaperone–client interaction strength, the folding time decreases first, and then increases. More detailed analysis showed that when the chaperone–client interaction is too strong, even small number of chaperones–client contacts can maintain the substrate bound with the chaperone. The sampling of the transient chaperones–client complex with sparse inter-molecule contacts makes the client protein have chance to access the misfolded state even it is bound with chaperone. The current results suggest that the interaction strength is an important factor controlling the Hsp70 chaperoning function.
