Protein recovery and identification from the gulf killifish, Fundulus grandis: Comparing snap-frozen and RNAlater® preserved tissues
Protein recovery and identification from the gulf killifish, Fundulus grandis: Comparing snap-frozen and RNAlater® preserved tissues
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DOI：
10.1002/pmic.201100328
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发表时间：
2011-11-01
期刊：
影响因子：
3.4
通讯作者：
Rees, Bernard B.
中科院分区：
文献类型：
--
作者：
Abbaraju, Naga V.;Cai, Yang;Rees, Bernard B.
可靠的生物组织蛋白质组学分析需要采样方法，以尽可能接近其体内状态保存蛋白质。在目前的研究中，评估了在液氮中快速冷冻组织或将新鲜组织浸泡在RNAlater(R)中后，墨西哥湾金枪鱼的五种组织的一维(1-D)凝胶中蛋白质丰度的模式。在肝脏和心脏中，一维凝胶中的蛋白质谱通过速冻保存得更好，而在鳃中，一维蛋白质谱通过浸泡在RNAlater(R)中保存得更好。在骨骼肌和大脑中，这两种方法产生了相似的蛋白质丰度模式。经LC-MS/MS分析和数据库检索，鉴定出肝脏中的17个蛋白质和鳃中的12个蛋白质。识别的蛋白质包括能量代谢酶、结构蛋白质和具有其他生物功能的蛋白质。这些对没有测序基因组的物种的蛋白质鉴定证明了大针茅作为脊椎动物环境蛋白质组学研究的模式生物的实用性。
Reliable proteomic analysis of biological tissues requires sampling approaches that preserve proteins as close to their in vivo state as possible. In the current study, the patterns of protein abundance in one-dimensional (1-D) gels were assessed for five tissues of the gulf killifish, Fundulus grandis, following snap-freezing tissues in liquid nitrogen or immersion of fresh tissues in RNAlater (R). In liver and heart, the protein profiles in 1-D gels were better preserved by snap-freezing, while in gill, the 1-D protein profile was better preserved by immersion in RNAlater (R). In skeletal muscle and brain, the two approaches yielded similar patterns of protein abundance. LC-MS/MS analyses and database searching resulted in the identification of 17 proteins in liver and 12 proteins in gill. Identified proteins include enzymes of energy metabolism, structural proteins, and proteins serving other biological functions. These protein identifications for a species without a sequenced genome demonstrate the utility of F. grandis as a model organism for environmental proteomic studies in vertebrates.