SIOC English
化学学报
高级检索

化学学报 ›› 2010, Vol. 68 ›› Issue (13): 1277-1284. 上一篇    下一篇

研究论文

分子模拟用于尤文肉瘤EWS-FLI1蛋白HLA-A2.1限制性CTL表位的筛选

曹凯*,1，黄路#,2，林治华3，舒勇1，韩智敏1，黄山虎1，廖翔1，王远强3，彭方毅3，安洪4   

  1. (1南昌大学第一附属医院 南昌 330006)
    (2江西省妇幼保健院 南昌 330006)
    (3重庆理工大学化学与生物工程学院 重庆 400050)
    (4重庆医科大学附属第一医院 重庆 400016)5. 重庆理工大学
    6. 重庆理工大学化学与生物工程学院
  • 投稿日期:2009-10-09 修回日期:2009-12-23 发布日期:2010-03-01
  • 通讯作者: 曹凯 E-mail:osteo.caokai@yahoo.com.cn
  • 基金资助:

    国家自然科学基金（30760254）；国家自然科学基金（60873103）

Screening of HLA-A2.1-Restricted CTL Epitopes Derived from Ewing s Sarcoma EWS-FLI1 Fusion Protein by Using Molecular Simulation

Cao Kai*,1 Huang Lu2 Lin Zhihua3 Shu Yong1 Han Zhimin1 Huang Shanhu1 Liao Xiang1, Wang Yuanqiang3 Peng Fangyi3 An Hong4   

  1. (1 The First Affiliated Hospital of Nanchang University, Nanchang 330006)
    (2Jiangxi Maternal and Child Health Hospital, Nanchang 330006)
    (3 Institute of Chemistry and Bioenginering, Chongqing University of Technology, Chongqing 400050)
    (4 The First Affiliated Hospital of Chongqing University of Medical Sciences, Chongqing 400016)
  • Received:2009-10-09 Revised:2009-12-23 Published:2010-03-01

PDF

1296

被引次数

6 | 2

为获得尤文肉瘤EWS-FLI1融合蛋白有效的HLA-A2.1限制性CTL表位, 综合运用BIMAS, SYFPEITHI, Predep和IEDB方案对EWS-FLI1进行CTL表位的预测, 得出8个CTL表位9肽序列; 应用多项式方案、量化基序方案对8个表位9肽进一步进行筛选, 获得其中4个9肽序列, 并进一步运用分子模拟方法初步模拟了4个表位肽与HLA-A2.1分子间的相互结合作用; 应用标准Fmoc方案合成CTL表位肽, RP-HPLC与MS分别鉴定合成肽的纯度与分子量; 通过亲和力实验验证了表位肽QIQLWQFLL(EWS-FLI1 304)与HLA-A2.1有较强的结合力, 从而为表位肽的后继免疫学特性研究提供了基础. 本研究提供了一个合理高效的表位筛选方法.

关键词: 尤文肉瘤, EWS-FLI1融合蛋白, CTL表位, 分子动力模拟

To obtain the HLA-A2.1-restricted CTL epitopes of EWS-FLI1 fusion protein of Ewing s sarcoma, EWS-FLI1 protein was synthetically predicted by BIMAS, SYFPEITHI, Predep and IEDB methods. Eight epitopes predicted by above methods were further screened by the polynomial method and the quantitative motif method as well. Four screened peptides were stimulated to dock with HLA-A2.1 by molecular dynamics simulation. All peptides were synthesized with standard Fmoc strategy, then detected beyond 98% in purity by reverse phase high performance liquid chromatography (RP-HPLC) and measured the values of molecular weight of them by mass spectrometry (MS). Infinity experiment validated the infinity of peptide QIQLWQFLL(EWS-FLI1 304) with HLA-A2.1 molecule, which could afford the foundation of immunological characteristics study of this screened peptide. This research provides a rational and high efficient method to screen epitope.

Key words: Ewing s sarcoma, EWS-FLI1 fusion protein, CTL epitope, molecular dynamics simulation