肺结核病人血清差异蛋白的分离及鉴定

Separation and identification of serum differential-expression proteins from pulmonary tuberculosis

目的分离并鉴定SELDI筛选出的肺结核病人血清差异表达蛋白。方法乙腈沉淀法去除血清中的高丰度蛋白,Tricine-SDS-PAGE电泳分离差异表达的目标蛋白,高效液相色谱串联质谱鉴定目标蛋白并进行生物信息学分析。结果电泳显示肺结核患者血清中小于6000Da的蛋白条带浓度明显低于正常人血清,经质谱鉴定和生物信息学分析,该目标蛋白为载脂蛋白(apo)C-I酶解片段,与SELDI建立的诊断模型中分子量为M4480.14的潜在标志物吻合。结论与血脂代谢密切相关的apo C-I可能是结核病血清潜在标志物之一。

Objective To separate and identify serum differential-expression proteins relevant to pulmonary tuberculosis（TB） screened by SELDI technology in previous studies.Methods The abundant high molecular weight proteins in serum from patients with active TB and controls were precipitated by acetonitrile,Tricine-SDS-PAGE were performed to separate serum target proteins,Target proteins were identified and analyzed by LC-MS/MS and bioinformatics respectively.Results The protein expression level of a band whose m/z value was smaller than 6.5 kDa was clearly lower than that in non-TB serum and was identified and analyzed as the protease digested fragment of apolipoprotein C-I（apo C-I）,in accordance with the target protein with a molecular weight similar to 4480.14 Da from the diagnostic model established by SELDI.Conclusion The protease digested fragment of apo C-I involved in lipoprotein metabolism may be one of potentially biomakers relevant to TB.