基于酶的自动化对角线毛细管电泳：在磷酸肽特征分析中的应用。
Automated enzyme-based diagonal capillary electrophoresis: application to phosphopeptide characterization.
机构信息
Department of Chemistry, University of Washington, Seattle, Washington 98195-1700, USA.
出版信息
Anal Chem. 2010 Feb 15;82(4):1564-7. doi: 10.1021/ac100029u.
Abstract
Diagonal capillary electrophoresis is a form of two-dimensional capillary electrophoresis that employs identical separation modes in each dimension. The distal end of the first capillary incorporates an enzyme-based microreactor. Analytes that are not modified by the reactor will have identical migration times in the two capillaries and will generate spots that fall on the diagonal in a reconstructed two-dimensional electropherogram. Analytes that undergo enzymatic modification in the reactor will have different migration times in the second capillary and will generate spots that fall off the diagonal in the electropherogram. We demonstrate the system with immobilized alkaline phosphatase to monitor the phosphorylation status of a mixture of peptides. This enzyme-based diagonal capillary electrophoresis assay appears to be generalizable; any post-translational modification can be detected as long as an immobilized enzyme is available that reacts with the modification under electrophoretic conditions.
摘要
对角线毛细管电泳是二维毛细管电泳的一种形式，它在每个维度中采用相同的分离模式。第一根毛细管的远端包含基于酶的微反应器。未被反应器修饰的分析物在两根毛细管中具有相同的迁移时间，并在重构的二维电泳图中生成落在对角线上的斑点。在反应器中经历酶修饰的分析物在第二根毛细管中的迁移时间不同，并在电泳图中生成偏离对角线的斑点。我们使用固定化碱性磷酸酶来演示该系统，以监测肽混合物的磷酸化状态。这种基于酶的对角线毛细管电泳分析似乎具有通用性；只要存在可在电泳条件下与修饰反应的固定化酶，就可以检测到任何翻译后修饰。
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