Plubell Deanna L, Wilmarth Phillip A, Zhao Yuqi, Fenton Alexandra M, Minnier Jessica, Reddy Ashok P, Klimek John, Yang Xia, David Larry L, Pamir Nathalie
From the ‡Department of Medicine, Knight Cardiovascular Institute, Oregon Health & Sciences University, Portland, Oregon.
§Proteomics Shared Resources, Oregon Health & Sciences University, Portland, Oregon.
Mol Cell Proteomics. 2017 May;16(5):873-890. doi: 10.1074/mcp.M116.065524. Epub 2017 Mar 21.
The lack of high-throughput methods to analyze the adipose tissue protein composition limits our understanding of the protein networks responsible for age and diet related metabolic response. We have developed an approach using multiple-dimension liquid chromatography tandem mass spectrometry and extended multiplexing (24 biological samples) with tandem mass tags (TMT) labeling to analyze proteomes of epididymal adipose tissues isolated from mice fed either low or high fat diet for a short or a long-term, and from mice that aged on low high fat diets. The peripheral metabolic health (as measured by body weight, adiposity, plasma fasting glucose, insulin, triglycerides, total cholesterol levels, and glucose and insulin tolerance tests) deteriorated with diet and advancing age, with long-term high fat diet exposure being the worst. In response to short-term high fat diet, 43 proteins representing lipid metabolism ( AACS, ACOX1, ACLY) and red-ox pathways ( CPD2, CYP2E, SOD3) were significantly altered (FDR < 10%). Long-term high fat diet significantly altered 55 proteins associated with immune response ( IGTB2, IFIT3, LGALS1) and rennin angiotensin system ( ENPEP, CMA1, CPA3, ANPEP). Age-related changes on low fat diet significantly altered only 18 proteins representing mainly urea cycle ( OTC, ARG1, CPS1), and amino acid biosynthesis ( GMT, AKR1C6). Surprisingly, high fat diet driven age-related changes culminated with alterations in 155 proteins involving primarily the urea cycle ( ARG1, CPS1), immune response/complement activation ( C3, C4b, C8, C9, CFB, CFH, FGA), extracellular remodeling ( EFEMP1, FBN1, FBN2, LTBP4, FERMT2, ECM1, EMILIN2, ITIH3) and apoptosis ( YAP1, HIP1, NDRG1, PRKCD, MUL1) pathways. Using our adipose tissue tailored approach we have identified both age-related and high fat diet specific proteomic signatures highlighting a pronounced involvement of arginine metabolism in response to advancing age, and branched chain amino acid metabolism in early response to high fat feeding. Data are available via ProteomeXchange with identifier PXD005953.
缺乏高通量方法来分析脂肪组织蛋白质组成限制了我们对负责与年龄和饮食相关的代谢反应的蛋白质网络的理解。我们开发了一种方法，使用多维液相色谱串联质谱法，并通过串联质谱标签（TMT）标记扩展多路复用（24个生物样品），以分析从短期或长期喂食低脂或高脂饮食的小鼠以及在低脂或高脂饮食下衰老的小鼠分离出的附睾脂肪组织的蛋白质组。外周代谢健康状况（通过体重、肥胖、血浆空腹血糖、胰岛素、甘油三酯、总胆固醇水平以及葡萄糖和胰岛素耐量试验来衡量）随着饮食和年龄增长而恶化，长期暴露于高脂饮食情况最差。对短期高脂饮食的反应中，43种代表脂质代谢（AACS、ACOX1、ACLY）和氧化还原途径（CPD2、CYP2E、SOD3）的蛋白质发生了显著变化（FDR<10%）。长期高脂饮食显著改变了55种与免疫反应（IGTB2、IFIT3、LGALS1）和肾素血管紧张素系统（ENPEP、CMA1、CPA3、ANPEP）相关的蛋白质。低脂饮食下与年龄相关的变化仅显著改变了18种主要代表尿素循环（OTC、ARG1、CPS1）和氨基酸生物合成（GMT、AKR1C6）的蛋白质。令人惊讶的是，高脂饮食驱动的与年龄相关的变化最终导致155种蛋白质发生改变，这些蛋白质主要涉及尿素循环（ARG1、CPS1）、免疫反应/补体激活（C3、C4b、C8、C9、CFB、CFH、FGA）、细胞外重塑（EFEMP1、FBN1、FBN2、LTBP4、FERMT2、ECM1、EMILIN2、ITIH3）和细胞凋亡（YAP1、HIP1、NDRG1、PRKCD、MUL1）途径。使用我们针对脂肪组织定制的方法，我们已经确定了与年龄相关和高脂饮食特异性的蛋白质组特征，突出了精氨酸代谢在应对年龄增长中的显著参与，以及支链氨基酸代谢在对高脂喂养早期反应中的参与。数据可通过ProteomeXchange获得，标识符为PXD005953。
