膜联蛋白AnxB1序列缺失突变体的结构模建及活性分析

为获得低分子量、低免疫原性的膜联蛋白AnxB1的序列缺失突变体 ,以AnxB1C端的 4个内部同源结构域为基础 ,模拟构建了 4个突变体群 .利用同源模建的方法对各突变体进行结构模建和分子优化 ,最后选择 4个结构最为合理的突变体在大肠杆菌GST融合表达系统中表达 .结果显示 :GST M3和GST M4均表达出较强的抗凝血活性 ,且免疫原性也降低为原来的 1 2 ,为进一步构建兼具抗凝、溶栓双重功能的靶向性溶栓药物奠定了基础 .

Molecular Modeling and Activity Analysis of Sequence-deleted Mutants of Annexin B1(AnxB1)

In order to obtain a low molecular weight and low immunogenicity mutant of annexin B1(AnxB1),four groups of mutants were constructed on the basis of the four internal homology domains of AnxB1. The tertiary structure of each mutant was established and optimized by homology modeling methods. Four of the optimized mutants were PCR amplified and inserted into pGEX 5T, and then expressed by induction with IPTG under the control of the tac promoter. The mutants GST M3, GST M4 kept the anticoagulatant activity and the immunogenicity decreased to a half of that in the wild GST AnxB1. The results provided the feasibility for further constructing a novel thromb targeted molecule which had both the anti coagulation and thrombolytic activity.