Protein profiling of human pancreatic islets by two-dimensional gel electrophoresis and mass spectrometry
Protein profiling of human pancreatic islets by two-dimensional gel electrophoresis and mass spectrometry
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DOI：
10.1021/pr050024a
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发表时间：
2005-05-01
影响因子：
4.4
通讯作者：
Bergsten, P
中科院分区：
文献类型：
--
作者：
Ahmed, M;Forsberg, J;Bergsten, P
人类基因组序列的完成为科学家提供了强大的资源来探索与糖尿病等疾病状态相关的分子事件。了解基因产物（尤其是蛋白质）的相对表达水平及其翻译后修饰至关重要。然而，尽管胰岛在葡萄糖稳态中发挥着关键作用，但人类的整体蛋白质表达数据却明显缺乏。我们在此报告人类胰岛的二维蛋白质图谱和数据库。凝胶中获得了高水平的重现性，总共检测到 744 个蛋白质点。我们已成功识别出对应 66 种不同蛋白质条目的 130 个点，并生成了人类胰岛的参考图谱。功能表征的蛋白质包括酶、分子伴侣、细胞结构蛋白、细胞防御蛋白、信号分子和转运蛋白。本研究中鉴定的许多蛋白质（例如，膜联蛋白 A2、延伸因子 1-α.2、组蛋白 H2B.a/g/k、热休克蛋白 90 beta、热休克 27 kDa 蛋白、亲环蛋白 B、过氧化还原蛋白 4；细胞角蛋白 7、18 和 19）以前在小鼠胰岛数据库中尚未描述。此外，多种蛋白质（如 GRP78、GRP94、PDI、钙网蛋白、膜联蛋白、细胞角蛋白、Profilin、热休克蛋白和 ORP150）表达的改变与糖尿病的发生有关。这项研究中提供的数据提供了人类胰岛蛋白质组的初稿参考图，这将为健康和糖尿病个体的胰岛的进一步蛋白质组分析铺平道路，从而深入了解这种情况的病理生理学。
Completion of the human genome sequence has provided scientists with powerful resources with which to explore the molecular events associated with disease states such as diabetes. Understanding the relative levels of expression of gene products, especially of proteins, and their post-translational modifications will be critical. However, though the pancreatic islets play a key role in glucose homeostasis, global protein expression data in human are decidedly lacking. We here report the two-dimensional protein map and database of human pancreatic islets. A high level of reproducibility was obtained among the gels and a total of 744 protein spots were detected. We have successfully identified 130 spots corresponding to 66 different protein entries and generated a reference map of human islets. The functionally characterized proteins include enzymes, chaperones, cellular structural proteins, cellular defense proteins, signaling molecules, and transport proteins. A number of proteins identified in this study (e.g., annexin A2, elongation factor 1-alpha. 2, histone H2B.a/g/k, heat shock protein 90 beta, heat shock 27 kDa protein, cyclophilin B, peroxiredoxin 4; cytokeratins 7, 18, and 19) have not been previously described in the database of mouse pancreatic islets. In addition, altered expression of several proteins, like GRP78, GRP94, PDI, calreticulin, annexin, cytokeratins, profilin, heat shock proteins, and ORP150 have been associated with the development of diabetes. The data presented in this study provides a first-draft reference map of the human islet proteome, that will pave the way for further proteome analysis of pancreatic islets in both healthy and diabetic individuals, generating insights into the pathophysiology of this condition.