双向凝胶电泳和质谱技术在睾丸蛋白表达研究中的应用

Application of Two-dimensional Electrophoresis and Mass Spectrometry in the Research of Testicular Proteins

目的 :探讨双向凝胶电泳和质谱技术在睾丸蛋白表达研究中的应用。 方法 :用双向凝胶电泳分离成年雄性ICR小鼠睾丸总蛋白 ;从胶上切下 2个蛋白点 ,经胶内原位酶解后 ,用质谱仪测得其肽质量指纹谱 ;再通过数据库检索鉴定蛋白质。 结果 :从胶上切下的 2个蛋白点数据库检索结果是小鼠血清白蛋白和蛋白质二硫化物异构酶。 结论 :用双向凝胶电泳分离睾丸蛋白样品取得了很高的分辨率 ,质谱技术鉴定蛋白快速方便 。

Objectives: To evaluate the application of two dimensional electrophoresis and mass spectrometry in the research of proteins expressed in testis. Methods: Protein from adult ICR mouse testes was extracted by two dimensional electrophoresis. Two protein spots were cut from the gel, then the proteins were digested in gel by enzyme and the generated peptides were measured by matrix assisted laser desorption/ionization time of flight mass spectrometry. The proteins were identified through database searching. Results: Two spots in Commasie Brilliant Blue stained gel were identified as serum albumin and protein disulfide isomerase by database searching. Conclusions: This rapid high resolution and efficient method is a very powerful way to analyze testicular proteins.