Proteomic characterization of the abdominal ganglion of Aplysia californica-A protein resource for neuroscience
Proteomic characterization of the abdominal ganglion of Aplysia californica-A protein resource for neuroscience
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DOI：
10.1002/pmic.201100641
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发表时间：
2012-08-01
期刊：
影响因子：
3.4
通讯作者：
Lubec, Gert
中科院分区：
文献类型：
--
作者：
Birner-Gruenberger, Ruth;Darnhofer, Barbara;Lubec, Gert
加州应用测试（AC）是一种广泛使用的学习和记忆测试模型。虽然已经产生了ESTs，但对AC蛋白的蛋白质组学研究有限。然而，在蛋白质水平上的研究是强制性的，这不仅是因为在核酸水平上的研究不能得出关于ptm的结论。因此，采用基于凝胶的蛋白质组学方法对AC的腹部神经节进行蛋白质分析。腹部神经节被切除，蛋白质被提取并在2DE上运行，随后用胰蛋白酶、凝乳胰蛋白酶和部分枯草菌素进行凝胶消化。采用纳米lc - esi - ltq - ft -质谱仪对多肽进行鉴定。通过检索NCBI非冗余公共AC EST数据库和NCBI非冗余公共AC protein数据库对MS/MS数据进行分析。共检测到363个蛋白质点代表的477种不同的蛋白质，并将其分配到不同的蛋白质途径，例如信号传导（受体、蛋白激酶和磷酸酶）、代谢、蛋白质合成、处理和降解、细胞骨架和结构、氧化氧化还原、热休克和伴侣、假设的、预测的和未命名的蛋白质。可溶性蛋白的蛋白质图谱的生成显示了迄今为止假设和预测的蛋白质的存在，并允许和挑战在蛋白质水平上的进一步工作，特别是在神经科学领域。
Aplysia californica (AC) is a widely used model for testing learning and memory. Although ESTs have been generated, proteomics studies on AC proteins are limited. Studies at the protein level, however, are mandatory, not only due to the fact that studies at the nucleic acid level are not allowing conclusions about PTMs. A gel-based proteomics method was therefore applied to carry out protein profiling in abdominal ganglia from AC. Abdominal ganglia were extirpated, proteins extracted and run on 2DE with subsequent in-gel digestion with trypsin, chymotrypsin, and partially by subtilisin. Peptides were identified using a nano-LC-ESI-LTQ-FT-mass spectrometer. MS/MS data were analyzed by searching the NCBI nonredundant public AC EST database and the NCBI nonredundant public AC protein database. A total of 477 different proteins represented by 363 protein spots were detected and were assigned to different protein pathways as for instance signaling (receptors, protein kinases, and phosphatases), metabolism, protein synthesis, handling and degradation, cytoskeleton and structural, oxido-redox, heat shock and chaperone, hypothetical, predicted and unnamed proteins. The generation of a protein map of soluble proteins shows the existence of so far hypothetical and predicted proteins and is allowing and challenging further work at the protein level, in particular in the field of neuroscience.