重组TRAIL的体外聚集形式及生物活性初步研究  

Pilot study on stability and activity of recombination TRAIL in vitro

作　　者：汤梅[1] 沈亚领[1] 周劲松 孙菁 球谊 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237 [2]上海歌佰德生物技术有限公司,上海200237

出　　处：《工业微生物》2013年第5期27-32,共6页Industrial Microbiology

基　　金：国家"重大新药创制"科技重大专项(2010ZX09401-301-3;2009ZX09103-748);上海市重点学科建设项目(B505);生物反应器工程国家重点实验室重点项目(No.2060204)资助

摘　　要：TRAIL(Tumor Related Apoptosis Inducing Ligand),又称凋亡素二配体(Apoptotin 2 Ligand),是肿瘤坏死因子(Tumour Necrosis Factor,INF)家族的成员,其重组可溶性表达的蛋白能诱导多种肿瘤细胞凋亡。通过分批发酵、纯化得到纯度在98%以上的可溶性重组TRAII。通过对环境pH以及Zn^(2+)对重组TRAIL的体外聚集形式影响的研究表明,pH能显著改变TRAIL的聚集形式,当环境pH从6.5降至5.0时,TRAIL主要以多聚体的形式存在。随着pH值的升高,TRAIL三聚体含量逐渐增多,当pH回升至6.5时,三聚体含量恢复至85%以上。Zn^(2+)影响实验表明,当经过透析实验使Zn^(2+)与TRAIL单体摩尔浓度比降至1:17时,TRAIL主要以单体形式存在,当重新升高体系Zn^(2+)浓度后,TRAIL又能恢复至三聚体形态。最后利用不同方式处理TRAIL的Zn^(2+)含量以得到不同三聚体含量的TRAIL,并对其进行活性测定实验,结果表明Zn^(2+)通过影响TRAIL的聚集形态可以显著影响TRAIL的生物活性。The effects of pH and Zn2＋ on monomer assembly forms of TRAIL were studied. Experiment results showed that pH could notably influence the aggregation of TRAIL. When pH decreased to 5.0, TRAIL existed as polymer in the solution. As pH increased, TRAIL existed as homotrimer. When pH was controlled at 6.5, the homotrimer of TRAIL were 85%. Experiments also revealed that when the mole ratio of Zn2＋ to TRAIL monomer decreased to 1 ： 17, most of the TRAIL existed as monomer forms. When the Zn2＋ concentration was increased, TRAIL could exist as homotrimer again. Ultimately, TRAIL with different contents of homotrimer was obtained by changing the Zn2＋ concentration. After the biologic activities were measured, the result indicated that Zn2＋ could affect bioactivity of TRAIL through influencing the homotrimer.

关 键 词：TRAIL PH Znn 三聚体 生物活性 

分 类 号：R730.2[医药卫生—肿瘤]