Kültz Dietmar, Li Johnathon, Paguio Darlene, Pham Tuan, Eidsaa Marius, Almaas Eivind
Department of Animal Science, University of California, Davis, Davis, CA 95616, USA.
Department of Animal Science, University of California, Davis, Davis, CA 95616, USA.
J Proteomics. 2016 Mar 1;135:112-131. doi: 10.1016/j.jprot.2015.10.002. Epub 2015 Oct 14.
Quantitative proteomics was used to reveal biochemical differences in kidneys of marine and freshwater three-spined sticklebacks. More than 1500 unambiguous proteins were identified, 106 of which are robustly co-translationally modified. Amino-terminal acetylation sites for 94 and proline hydroxylation sites for 12 proteins, including 4 protein disulfide isomerases having the consensus motif APWCGHCK, were determined. More than 1500 proteins were quantified by LC-MS/MS yielding 120 proteins with consistent population-specific abundance differences. Twenty-five of these were selected for validation by data-independent acquisition (DIA) and spectral library based MS2 quantitation. A dense biochemical network was revealed, which promotes the synthesis of the organic osmolytes betaine, sorbitol, trimethylamine oxid (TMAO), and urea. It contains 33 of 49 proteins that are elevated in marine compared to freshwater sticklebacks, including the most highly elevated proteins (dimethylaniline monooxygenase, alanine-glyoxylate aminotransferase, glycine N-methyltransferase). Freshwater stickleback kidneys contain elevated levels of proteolytic, cytoskeletal, extracellular matrix, and calcium signaling proteins. Proteins that are most elevated in freshwater sticklebacks are ES1 protein homolog, apoptosis-associated speck-like protein containing a CARD and caspase 1. Protein-abundance network analysis demonstrates significantly higher levels of synchronized abundance control in marine sticklebacks. The significance of these findings for biochemical diversification of renal function in marine and FW sticklebacks is discussed.
定量蛋白质组学被用于揭示海洋和淡水三刺鱼肾脏中的生化差异。共鉴定出1500多种明确的蛋白质，其中106种有强烈的共翻译修饰。确定了94种蛋白质的氨基末端乙酰化位点和12种蛋白质的脯氨酸羟化位点，其中包括4种具有共有基序APWCGHCK的蛋白质二硫键异构酶。通过液相色谱-串联质谱法对1500多种蛋白质进行了定量分析，得到了120种具有一致的种群特异性丰度差异的蛋白质。从中选择了25种通过数据非依赖采集（DIA）和基于谱图库的MS2定量进行验证。揭示了一个密集的生化网络，该网络促进有机渗透剂甜菜碱、山梨醇、三甲胺氧化物（TMAO）和尿素的合成。与淡水三刺鱼相比，该网络包含49种蛋白质中的33种在海洋三刺鱼中含量升高的蛋白质，包括含量升高最高的蛋白质（二甲基苯胺单加氧酶、丙氨酸-乙醛酸转氨酶、甘氨酸N-甲基转移酶）。淡水三刺鱼肾脏中蛋白水解、细胞骨架、细胞外基质和钙信号蛋白的水平升高。在淡水三刺鱼中含量升高最高的蛋白质是ES1蛋白同源物、含有CARD的凋亡相关斑点样蛋白和半胱天冬酶1。蛋白质丰度网络分析表明，海洋三刺鱼中同步丰度控制水平显著更高。讨论了这些发现对海洋和淡水三刺鱼肾功能生化多样化的意义。
