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脂质筏的 Flot2 成分在 P19C6 细胞的神经分化过程中改变定位。

The Flot2 component of the lipid raft changes localization during neural differentiation of P19C6 cells.

机构信息

School of Life Science and Technology, Tokyo Institute of Technology, Meguro-ku, Tokyo, Japan.

出版信息

BMC Mol Cell Biol. 2019 Aug 27;20(1):38. doi: 10.1186/s12860-019-0225-0.

Abstract

BACKGROUND

Flotillin-2 (Flot2) is a lipid raft scaffold protein that is thought to be related to neural differentiation. Flot2 is phosphorylated by Fyn, a Src kinase, and causes raft-dependent endocytosis; however, the exact role of Flot2 in neural differentiation remains unclear. To reveal the roles of lipid raft-associated proteins during neural differentiation, we tried to analyze the expression and localization.

RESULTS

In this study, we found that the expression levels of the Flot2 and Fyn proteins increased in whole-cell lysates of P19C6 cells after neural differentiation. In addition, sucrose density fractionation and immunofluorescence experiments revealed an increase in the localization of Flot2 and Fyn to lipid rafts after neural differentiation. We also found that Fyn partially colocalized with Flot2 lipid rafts in neural cells.

CONCLUSION

The observed distribution of Fyn and level of inactivated Fyn and/or c-Src in detergent-resistant membrane (DRM) fractions suggests that the amount of activated Fyn might increase in DRM fractions after neural differentiation. Overall these findings suggest that Flot2 lipid rafts are associated with Fyn, and that Fyn phosphorylates Flot2 during neural differentiation of P19C6 cells.

摘要

背景

Flotillin-2(Flot2)是一种脂质筏支架蛋白,被认为与神经分化有关。Flot2 被Src 激酶 Fyn 磷酸化,并导致筏依赖的内吞作用;然而,Flot2 在神经分化中的确切作用仍不清楚。为了揭示脂质筏相关蛋白在神经分化过程中的作用,我们试图分析其表达和定位。

结果

在这项研究中,我们发现 P19C6 细胞在神经分化后,全细胞裂解物中 Flot2 和 Fyn 蛋白的表达水平增加。此外,蔗糖密度梯度分离和免疫荧光实验显示,神经分化后 Flot2 和 Fyn 向脂质筏的定位增加。我们还发现 Fyn 部分与神经细胞中的 Flot2 脂质筏共定位。

结论

观察到 Fyn 和失活的 Fyn 和/或 c-Src 在去污剂抗性膜(DRM)部分的分布表明,在神经分化后,DRM 部分中激活的 Fyn 可能增加。总的来说,这些发现表明 Flot2 脂质筏与 Fyn 相关,并且在 P19C6 细胞的神经分化过程中,Fyn 磷酸化 Flot2。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/364d/6712619/d612d1243105/12860_2019_225_Fig1_HTML.jpg

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