Proteomic analysis of murine kidney proximal tubule sub-segment derived cell lines reveals preferences in mitochondrial pathway activity.
Proteomic analysis of murine kidney proximal tubule sub-segment derived cell lines reveals preferences in mitochondrial pathway activity.
复制标题
对小鼠肾近端小管亚段衍生细胞系的蛋白质组学分析揭示了线粒体途径活性的偏好。
DOI：
10.1016/j.jprot.2023.104998
复制
发表时间：
2023
影响因子：
3.3
通讯作者：
Bacallao,RobertL
中科院分区：
文献类型：
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作者：
Ferreira,RicardoMelo;deAlmeida,Rita;Culp,Clayton;Witzmann,Frank;Wang,Mu;Kher,Rajesh;Nagami,GlennT;Mohallem,Rodrigo;Andolino,ChaylenJade;Aryal,UmaK;Eadon,MichaelT;Bacallao,RobertL
近端小管 (PT) 是肾单位的一部分，负责肾脏中大部分溶质和水的重吸收。它的每个子部分都有专门的功能；然而，对于决定 PT 亚片段氧化磷酸化能力的基因和蛋白质知之甚少。这些信息对于了解肾功能至关重要，并将提供肾细胞对损伤、生理应激源和发育的适应的全面概况。本研究分析了三种永生化鼠肾细胞系（PT S1、S2 和 S3 片段）的蛋白质含量，并将它们与鼠成纤维细胞系进行比较。所有三种近端小管细胞系主要通过氧化磷酸化产生 ATP，而成纤维细胞系则进行糖酵解。蛋白质组数据表明，细胞系之间蛋白质组特征最显着的差异是已知位于细胞核中的蛋白质，其次是线粒体蛋白质。使用近端小管细胞系进行线粒体代谢底物利用测定，以确定底物利用动力学，从而为蛋白质组数据集提供生理背景。这些数据将使研究人员能够研究肾单位特异性细胞系、上皮细胞和成纤维细胞之间以及活跃呼吸细胞和糖酵解细胞之间的差异。意义将永生化小鼠肾近曲小管细胞中表达的蛋白质与小鼠成纤维细胞系蛋白质组进行比较。近端小管段特异性细胞系：S1、S2和S3均在细胞通过氧化磷酸化产生ATP而成纤维细胞系利用无氧糖酵解产生ATP的条件下生长。蛋白质组学研究允许以下查询：1) 近端小管段特异性细胞系之间的比较，2) 极化上皮细胞和成纤维细胞之间的比较，3) 采用氧化磷酸化与无氧糖酵解的细胞之间的比较，以及 4) 在透明与不透明膜支持物上生长的细胞之间的比较。数据发现核蛋白表达和线粒体蛋白存在主要差异。对于需要永生化肾近曲小管上皮细胞进行研究的研究人员来说，该蛋白质组数据集将是重要的基线数据集。
The proximal tubule (PT) is a nephron segment that is responsible for the majority of solute and water reabsorption in the kidney. Each of its sub-segments have specialized functions; however, little is known about the genes and proteins that determine the oxidative phosphorylation capacity of the PT sub-segments. This information is critical to understanding kidney function and will provide a comprehensive landscape of renal cell adaptations to injury, physiologic stressors, and development. This study analyzed three immortalized murine renal cell lines (PT S1, S2, and S3 segments) for protein content and compared them to a murine fibroblast cell line. All three proximal tubule cell lines generate ATP predominantly by oxidative phosphorylation while the fibroblast cell line is glycolytic. The proteomic data demonstrates that the most significant difference in proteomic signatures between the cell lines are proteins known to be localized in the nucleus followed by mitochondrial proteins. Mitochondrial metabolic substrate utilization assays were performed using the proximal tubule cell lines to determine substrate utilization kinetics thereby providing a physiologic context to the proteomic dataset. This data will allow researchers to study differences in nephron-specific cell lines, between epithelial and fibroblast cells, and between actively respiring cells and glycolytic cells.SignificanceProteomic analysis of proteins expressed in immortalized murine renal proximal tubule cells was compared to a murine fibroblast cell line proteome. The proximal tubule segment specific cell lines: S1, S2 and S3 are all grown under conditions whereby the cells generate ATP by oxidative phosphorylation while the fibroblast cell line utilizes anaerobic glycolysis for ATP generation. The proteomic studies allow for the following queries: 1) comparisons between the proximal tubule segment specific cell lines, 2) comparisons between polarized epithelia and fibroblasts, 3) comparison between cells employing oxidative phosphorylation versus anaerobic glycolysis and 4) comparisons between cells grown on clear versus opaque membrane supports. The data finds major differences in nuclear protein expression and mitochondrial proteins. This proteomic data set will be an important baseline dataset for investigators who need immortalized renal proximal tubule epithelial cells for their research.