中华皮肤科杂志 ›› 2007, Vol. 40 ›› Issue (1): 48-50.

• 论著 • 上一篇    下一篇

蛋白质组学技术鉴定白癜风相关的黑素细胞膜抗原

李强1, 高天文1, 焦彬2, 于晓云1, 胡雪慧1, 卢涛3, 栾琪1, 刘玲1, 刘玉峰1   

  1. 1. 第四军医大学西京医院皮肤科 西安 710032;
    2. 北京武警总队医院皮肤科;
    3. 武警医学院附属医院皮肤科
  • 收稿日期:2006-02-20 出版日期:2007-01-15 发布日期:2007-01-15
  • 通讯作者: 高天文,email:gaotw@fmmu.edu.cn E-mail:gaotw@fmmu.edu.cn
  • 作者简介:李强,email:qiangli@fmmu.edu,cn

Identification of melanocgte surface antigens in patients with vitiligo by proteomic analysis

LI Qiang1, GAO Tian-wen1, JIAO Bin2, YU Xiao yun1, HU Xue-hui1, LU Tao3, LUAN Qi1, LIU Ling1, LIU Yu-feng1   

  1. Department of Dermatology, Xijing Hospital, the Fourth Military Medical University, Xi'an 710032, China
  • Received:2006-02-20 Online:2007-01-15 Published:2007-01-15

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摘要： 目的 通过蛋白质组学技术鉴定白癜风自身抗体识别的黑素细胞膜抗原的性质.方法 采用活细胞ELISA和免疫印迹法筛选含高滴度抗黑素细胞抗体的白癜风患者血清.提取黑素细胞膜表面蛋白,制备2块平行级双向电泳凝胶,免疫印迹,比较健康对照血清与白癜风患者血清免疫着色差异点,通过图像分析及蛋白质组学服务器查询、分析差异蛋白的性质.结果 黑素细胞膜蛋白的双向电泳显示500个左右的蛋白点,分离效果理想.双向电泳的免疫印迹成功发现3个蛋白差异点,其中一个为已知的细胞膜抗原黑素细胞浓集受体-1(MCHR-1),另两个蛋白点的表观相对分子质量约为75001)和60000,表观等电点约为6.5和5.5,无法通过生物信息学技术获得其性质,有待进一步进行质谱鉴定.结论 通过蛋白质学技术初步鉴定出3个位于黑素细胞膜的白癜风候选自身抗原,一个是已知的膜抗原MCHR-1.另外2个还需进一步鉴定.

关键词: 白癜风, 蛋白质组学, 黑素细胞, 膜蛋白质类, 自身抗原

Abstract: Objective To identify the melanocyte surface antigens associated with vitiligo by a proteomic approach. Methods Sera were screened for the presence of a high titer of antibodies against melanocytes from 256 patients with vitiligo by cellular enzyme-linked immunosorbent assay(cellular ELISA)and Western blotting. The membrane proteins of melanocytes were extracted and separated by two-dimensional gel electrophoresis(2-DE).Separated membrane proteins were transferred to polyvinylidene difluoride(PVDF)membrane and immunoblotted with either pooled sera of 10 patients with vitiligo or those of healthy controls. The different spots were analyzed by image analysis software, followed by identification using the bioinformatics tool of ExPASy(Expert Protein Analysis System)proteomics server. Results About 500 distinct protein spots were identified by 2-DE, of which 3 spots were unique to the sera of patients with vitiligo. One of them was identified as melanin-concentrating hormone receptor 1(MCHR-1), a known autoantigen of vitiligo. The other two proteins could not be identified using the bioinformatics tool and might need to be identified by mass spectrometry. The apparent molecular weight and isoelectric point of one protein were determined to be 75 000 and 6.5, whereas those of another one were 60 000 and 5.5, respectively. Conclusion Three melanocyte surface antigens are identified as candidate autoantigens in vitilieo by nroteomic analysis. One of them is MCHR-1, and the other two remain to be identified.

Key words: Vitiligo, Proteomics, Melanocytes, Membrane proteins, Autoantigens