可溶性人类白细胞抗原F和分化簇8α同二聚体的表达、纯化与相互作用

Expression,purification and interaction of human leukocyte antigen F and cluster of differentiation 8α homodimers

为了获得大量可溶性人类白细胞抗原F(Human leukocyte antigen F,HLA-F)和分化簇8α同二聚体(Cluster of differentiation 8αhomodimers,CD8αα)蛋白并对它们的相互关系进行研究,通过同义突变的方法改变了HLA-F和CD8αα基因序列N端的大肠杆菌稀有密码子,获得了高效表达的HLA-F和CD8αα包涵体蛋白;所表达的蛋白通过稀释法复性后,分别进行了凝胶过滤层析和离子交换纯化。经凝胶过滤层析和native-PAGE检测,推测HLA-F与CD8αα间具有相互作用。该相互作用可能影响其他MHC分子与CD8αα结合,从而调节免疫反应。研究结果对了解和进一步深入研究HLA-F的功能提供了线索。

To obtain large quantity of human leukocyte antigen F（HLA-F） and cluster of differentiation 8α homodimers（CD8αα） proteins and to study their relationship,HLA-F and CD8α genes with rare codon in Escherichia coli were cloned using an N-terminal synonymous mutation method.High-efficiency expression protein inclusion bodies were acquired.The proteins were refolded using the dilution method and purified with gel-filtration and anion exchange chromatography.The results of gel-filtration and native-PAGE indicate that HLA-F interacts with CD8αα.This interaction may affect the binding between CD8αα and other MHC molecules to regulate immune responses.These results provide a basis for further research of HLA-F.