Zhang Bo, Wang Ren-Zhi, Fan Ming, Yao Yong, Li Gui-Lin
Department of Neurosurgery, First Affiliated Hospital, Dalian Medical University, Dalian 116011, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2006 Feb;22(1):26-9.
To initially analyze the protein expression in the rat hippocampus with the proteomics approach.
Proteins from hippocampal tissue homogenates of the rat were separated by two-dimensional gel electrophoresis(2-DE), and the proteins were stained with colloidal coomassie blue to produce a high-resolution map of the rat hippocampus proteome. Selected proteins from this map were digested with trypsin, and the resulting tryptic peptides were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The mass spectrometric data were used to identify the proteins through searches of the NCBI protein sequence database.
37 prominent proteins with various functional characteristics were identified. The identification of brain protein classes, like metabolism enzymes, cytoskeleton proteins, heat shock proteins, antioxidant proteins, signalling proteins, proteasome-related proteins, neuron-specific proteins and glial-associated proteins. Furthermore, 3 hypothetical proteins which are unknown proteins, so far only proposed from their nucleic acid structure were identified.
This study provides the first unbiased characterization of proteins of the rat hippocampus and will be used for future studies of differential protein expression in rat models of neurological disorders.
采用蛋白质组学方法初步分析大鼠海马体中的蛋白质表达情况。
大鼠海马组织匀浆中的蛋白质通过二维凝胶电泳(2-DE)进行分离,并用考马斯亮蓝胶体染色,以生成大鼠海马蛋白质组的高分辨率图谱。从该图谱中选取的蛋白质用胰蛋白酶消化,所得的胰蛋白酶肽段通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)进行分析。质谱数据用于通过搜索NCBI蛋白质序列数据库来鉴定蛋白质。
鉴定出37种具有不同功能特征的显著蛋白质。鉴定出了脑蛋白类别,如代谢酶、细胞骨架蛋白、热休克蛋白、抗氧化蛋白、信号蛋白、蛋白酶体相关蛋白、神经元特异性蛋白和神经胶质相关蛋白。此外,还鉴定出3种假设蛋白,它们是未知蛋白,目前仅根据其核酸结构提出。
本研究首次对大鼠海马体蛋白质进行了无偏性表征,并将用于未来神经疾病大鼠模型中差异蛋白质表达的研究。