Protein carbonhydrate Analysis: gel-based staining, liquid chromatography, mass spectrometry, and microarray screening.
Protein carbonhydrate Analysis: gel-based staining, liquid chromatography, mass spectrometry, and microarray screening.
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蛋白质碳水化合物分析：凝胶染色、液相色谱、质谱和微阵列筛选。
DOI：
10.1007/978-1-60327-047-2_2
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
Ye,Bin
中科院分区：
文献类型：
--
作者：
Shin,Injae;Zamfir,AlinaD;Ye,Bin
碳水化合物对蛋白质和脂质的修饰在功能上“微调”这些分子，这些分子通过特定的生物相互作用参与了各种各样的生理和致病过程。因此，糖基化错误具有严重的影响，并与许多常见疾病如癌症和糖尿病有关。碳水化合物历来是药物发明的理想目标。碳水化合物检测和分析的最新进展促进了对糖蛋白和碳水化合物在许多细胞和信号过程中的作用的理解，为碳水化合物在疾病诊断和药物发现中的利用铺平了道路。在这里，我们介绍了最新开发的分析方法，这些方法已在许多研究实验室中常规使用。在通过一维（1D）和二维（2D）聚丙烯酰胺凝胶电泳（PAGE）分离蛋白质之后，凝胶染色方法提供了鉴定候选糖蛋白的第一步，以便进行进一步的表征。液相色谱和质谱联用为糖蛋白（包括糖肽和聚糖）的定量分析和糖基化位点的鉴定提供了宝贵的技术。基于碳水化合物的微阵列系统的先进技术结合了高通量分析，扩展了碳水化合物介导的分子相互作用的生物医学研究范围。
Carbohydrate modification of proteins and lipids serves to functionally “fine-tune” these molecules, which are involved in a wide variety of physiological and pathogenic processes through specific biological interactions. Therefore, errors in glycosylation have severe implications and have been associated with many common diseases such as cancer and diabetes. Carbohydrates have historically been desirable targets for drug invention. Recent advances in detection and analysis of carbohydrates have facilitated understanding of the roles of glycoproteins and carbohydrates in many cellular and signaling processes, paving the way for the exploitation of carbohydrates in disease diagnosis and drug discovery. Here, we introduce the most recently developed analytical methods, which have been routinely used in many research laboratories. Following the protein separation by one-dimensional (1D) and two-dimensional (2D) polyacrylamide gel electrophoresis (PAGE), gel-based staining methods provide the initial step to identify candidate glycoproteins so that further characterization can proceed. Combined liquid chromatography and mass spectrometry provide an invaluable technology for quantitative analysis of glycoproteins including glycopeptides and glycans and to identify the glycosylation site. The advanced technology of the carbohydrate-based microarray system incorporates high-throughput analysis to extend the scope of biomedical research on carbohydrate-mediated molecular interactions.