Characterization of the sebocyte lipid droplet proteome reveals novel potential regulators of sebaceous lipogenesis
Characterization of the sebocyte lipid droplet proteome reveals novel potential regulators of sebaceous lipogenesis
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DOI：
10.1016/j.yexcr.2014.12.004
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发表时间：
2015-03-01
影响因子：
3.7
通讯作者：
Schneider, Marlon R.
中科院分区：
文献类型：
--
作者：
Dahlhoff, Maik;Froehlich, Thomas;Schneider, Marlon R.
脂质代谢依赖于脂滴（LD），脂滴是由富含蛋白质的磷脂单层包围的细胞质结构。虽然脂质合成是皮脂腺细胞分化的标志，但皮脂腺细胞的LD相关蛋白尚未得到系统的评价，通过密度梯度离心收集SZ 95皮脂腺细胞的LD组分，并通过纳米液体色谱/串联质谱分析相关蛋白。其中54个蛋白质在LD组分中显著富集，其中6个蛋白质在LD组分中未检测到。LD组分含有高水平的典型LD相关蛋白，如PLIN 2/PLIN 3，并且大多数蛋白属于LD相关蛋白的功能类别，表明数据集可靠。在通过gRT-PCR确认编码六种先前未鉴定的蛋白质的转录物在SZ 95皮脂腺细胞和另一种皮脂腺细胞系（SebE 6 E7）中的表达后，我们专注于其中两种蛋白质，ALDH 1A 3和EPHX 4。虽然EPHX 4几乎完全位于LD的表面上，但ALDH 1A 3显示出更广泛的定位，包括额外的细胞质结构。siRNA介导的下调显示EPHX 4的缺失增加LD大小和皮脂腺脂肪生成。对这些蛋白在皮脂细胞生理学和皮脂腺脂肪生成中的作用的进一步研究可能为治疗皮脂腺相关疾病如痤疮提供新的策略。(C)2014爱思唯尔公司All rights reserved.
Lipid metabolism depends on lipid droplets (LD), cytoplasmic structures surrounded by a proteinrich phospholipid monolayer. Although lipid synthesis is the hallmark of sebaceous gland cell differentiation, the LD-associated proteins of sebocytes have not been evaluated systematically.The LD fraction of SZ95 sebocytes was collected by density gradient centrifugation and associated proteins were analyzed by nanoliquid chromatography/tandem mass spectrometry. 54 proteins were significantly enriched in LD fractions, and 6 of them have not been detected previously in LDs. LD fractions contained high levels of typical LD-associated proteins as PLIN2/PLIN3, and most proteins belonged to functional categories characteristic for LD-associated proteins, indicating a reliable dataset. After confirming expression of transcripts encoding the six previously unidentified proteins by gRT-PCR in SZ95 sebocytes and in another sebocyte line (SebE6E7), we focused on two of these proteins, ALDH1A3 and EPHX4. While EPHX4 was localized almost exclusively on the surface of LDs, ALDH1A3 showed a more widespread localization that included additional cytoplasmic structures. siRNA-mediated downregulation revealed that depletion of EPHX4 increases LD size and sebaceous lipogenesis. Further studies on the roles of these proteins in sebocyte physiology and sebaceous lipogenesis may indicate novel strategies for the therapy of sebaceous gland-associated diseases such as acne. (C) 2014 Elsevier Inc. All rights reserved.