摘要
【目的】应用二维电泳和质谱对正常人眼小梁组织蛋白质组的进行初步分析。【方法】采用二维凝胶电泳对正常人眼小梁组织进行蛋白分离和图像分析。分别用(7cm×8cm、13cm×16cm、18cm×16cm)3种不同尺寸大小凝胶分离蛋白,并结合基质辅助激光解吸附电离飞行时间质谱分析及数据库搜索从而鉴定部分蛋白质斑点。【结果】建立小梁组织蛋白样品处理及二维凝胶电泳的实验条件及方法,获得正常人眼小梁组织二维凝胶电泳蛋白图谱。3种不同尺寸大小凝胶的蛋白质分离斑点数分别为230个、875个、1213个。质谱鉴定出14个蛋白点。【结论】完成人眼小梁组织蛋白质的二维电泳凝胶“标准”图谱并作质谱分析的方法性探索对同类研究具有参考意义,为分析小梁组织在青光眼发病过程中蛋白质表达改变研究提供了新的方法及途径。
[Objective] To investigate the proteome of normal human trabecular meshwork （TM） by twodemensional gel electrophoresis（2-DE） and mass spectrometry. [Methods] Protein of the normal human TM was separated by two-demensional gel electrophoresis at various gel sizes （7 cm×8 cm, 13 cm×16 cm, 18 cm×16 cm）. Image analysis was carried out with Image Master Elite 3.10 software package. Protein spots on 2-DE gel were identified by matrix assisted laser desorption/ionization-time-of-flight mass pectrometry （MALDI-TOF-MS） and database searching. [Results] Systemic processing and essential conditions of sample preparation and 2-DE for human trabecular meshwork were established successfully.The 2-DE gel patterns of human trabecular meshwork with three sizes （7 cm×8 cm,13 cm×16 cm,18 cm×16 cm）were obtained.There were about 230,875 and 1213 protein spots detected on these three gels respectively. 14 protein spots were identified successfully by MALDI-TOF-MS. [ Conclusion ] The approach of proteomic analysis using 2-DE and MALDI-TOF-MS and the gel patterns of human TM were accomplished successfully, which offers a new approach for further study on human TM protein related glaucoma.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2006年第6期672-676,共5页
Journal of Sun Yat-Sen University:Medical Sciences
基金
国家自然科学基金资助项目(30371513)
广东省重大科技专项(2004A30801001)
广东省自然科学基金资助项目(031732)
关键词
小梁组织
二维凝胶电泳
质谱
蛋白图谱
trabecular meshwork
two-demensional gel electrophoresis
mass spectrometry
protein map
