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膜联蛋白AnxB1是我们发现的一个新的膜联蛋白亚家族成员,具较强的抗凝血活性和血栓亲和性.为更深入研究AnxB1的结构与功能,首先利用同源建模的方法预测AnxB1的三维结构,发现AnxB1具有四个内部同源结构域,其中1,2,4结构域的Cα主链叠合非常一致.以此为基础,构建四个序列缺失突变体M1,M2,M3,M4,并在大肠杆菌GST融合表达系统表达,其中的两个突变体GST-M3,GST-M4不仅保留了较强的抗凝血活性,而且免疫原性大大降低,分子量也分别减少到27和34kD.为进一步构建高效、低免疫原性的靶向性溶栓药物奠定了基础.
Annexin AnxB1 is a new member of Annexin subfamily that we found, with strong anticoagulant activity and thrombus affinity.In order to further study the structure and function of AnxB1, we first used the method of homology modeling The three-dimensional structure of AnxB1 was predicted and AnxB1 was found to have four internal homology domains, in which the Cα backbones of 1, 2 and 4 domains were very consistent.On the basis of this, four sequence deletion mutants M1, M2, M3 and M4 were expressed in Escherichia coli GST fusion expression system. The two mutants GST-M3 and GST-M4 not only retained strong anticoagulant activity, but also greatly reduced immunogenicity and molecular weight to 27 and 34kD, which laid the foundation for the further construction of efficient and low immunogenic targeting thrombolytic drugs.