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

摘要:

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 8α homodimers，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-

Abstract:

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.