Lipin1, a member of the lipin family, serves as a phospholipid phosphatase or a co-transcriptional regulator in lipid metabolism. Recent studies also show that lipin1 is involved in many other cellular metabolism processes. However, the clear regulatory mechanism for lipin1 is unknown. The 293T human renal epithelial cell line represents a commonly used and well established expression system for recombinant proteins. Herein, we used two-dimensional polyacrylamide gel electrophoresis (2D-GE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to explore the changes in protein expression induced by lipin1 overexpression in 293T cells. Western blotting was used to confirm one of the expression changes of related proteins. Subsequently, the function and relationship of these proteins were analyzed by bioinformatics approach. By using 2D-PAGE, approximately 152 proteins were separated and eleven proteins were found to be significantly affected by lipin1 overexpression compared to the control. Among them, three proteins (eEF-1B γ, CCT1 and CCT3) were up-regulated and other eight proteins (NDKA, Stathmin, HNRNP A1, TK, KRT1, PKM, RanBP1 and LDHB) were down-regulated. These proteins were successfully identified with peptide mass fingerprinting using MALDI-TOF-MS after in-gel trypsin digestion. The bioinformatic analysis showed that these proteins are classified into seven protein species, including transferase, cleavage enzyme, cytoskeleton protein, chaperone protein, regulatory protein, structural protein and oxidoreductase. The results highlight the potential roles of lipin1 involved in many cellular metabolism processes, including myelin synthesis, extracellular domain formation, membrane bound vesicle synthesis and companion protein T complex synthesis.

中文翻译：

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293T人肾上皮细胞对lipin1过表达的蛋白质组学变化

脂蛋白家族的一个成员，脂蛋白1在脂质代谢中充当磷脂磷酸酶或共转录调节剂。最近的研究还表明，lipin1参与许多其他细胞代谢过程。但是，尚不清楚lipin1的明确调控机制。293T人肾上皮细胞系代表了重组蛋白的一种常用且建立良好的表达系统。本文中，我们使用二维聚丙烯酰胺凝胶电泳（2D-GE）和基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS）来研究293T中lipin1过表达诱导的蛋白质表达变化。细胞。使用蛋白质印迹法来证实相关蛋白的表达变化之一。后来，通过生物信息学方法分析了这些蛋白质的功能和相互关系。通过使用2D-PAGE，与对照相比，大约152种蛋白质被分离，并且发现11种蛋白质受到lipin1过表达的显着影响。其中，三种蛋白（eEF-1Bγ，CCT1和CCT3）被上调，而其他八种蛋白（NDKA，Stathmin，HNRNP A1，TK，KRT1，PKM，RanBP1和LDHB）被下调。凝胶内胰蛋白酶消化后，使用MALDI-TOF-MS通过肽质量指纹图谱成功鉴定了这些蛋白质。生物信息学分析表明，这些蛋白被分为七个蛋白种类，包括转移酶，切割酶，细胞骨架蛋白，伴侣蛋白，调节蛋白，结构蛋白和氧化还原酶。