表面等离子共振法测量乙肝病毒功能受体NTCP与配体相互作用  

Study on the interaction between hepatitis B virus functional receptor NTCP and ligands by surface plasmon resonance

作　　者：覃覃 曾苏[1] QIN Tan;ZENG Su(Institute of Drug Metabolism and Pharmaceutical Analysis,College of Pharmaceutical Sciences,Zhejiang University,Hangzhou 310058,China)

机构地区：[1]药物代谢和药物分析研究所,浙江大学药学院,杭州310058

出　　处：《分析试验室》2022年第9期999-1005,共7页Chinese Journal of Analysis Laboratory

基　　金：国家自然科学基金项目(81903705);中央高校基本科研业务费专项资助。

摘　　要：钠离子/牛磺胆酸共转运多肽(NTCP,SLC10A1)是乙型肝炎病毒(HBV)感染肝细胞的功能性受体,也是抗慢性乙肝感染药物的潜在靶标。目前评价NTCP与其配体相互作用的方法仍然有限。本文使用Sf9细胞表达并纯化了重组NTCP蛋白,通过抗体间接偶联的方法将其固定在表面等离子共振实验芯片上。通过动力学分析,拟合得出HBV PreS1多肽对NTCP结合的平衡解离常数K_(D)为4.93μmol/L;NTCP经典底物牛磺胆酸钠对NTCP的平衡解离常数K为25.3μmol/L。相对于化合物抑制NTCP底物转运的活性,通过SPR方法测得的平衡解离常数可比较化合物与NTCP直接相互作用力的强弱,并用于研发基于NTCP-PreS1相互作用的抗HBV药物。Sodium/taurocholate cotransporting polypeptide(NTCP,SLC10A1)mediates the entry of hepatitis B virus(HBV)into hepatocytes.NTCP is the functional receptor for HBV infection and a potential target for anti-HBV drugs.At present,methods that evaluate the interaction between NTCP and its ligands are still limited.In this paper,the recombinant NTCP was expressed by Sf9 cells and purified,and fixed on the surface plasmon resonance(SPR)sensor chip by indirect antibody coupling.According to SPR kinetic analysis,the equilibrium dissociation constant K_(D)of HBV PreS1 polypeptide binding to NTCP is 4.93μmol/L.The equilibrium dissociation constant K_(D)of classic substrate taurocholate sodium for NTCP is 25.3μmol/L.Rather than the inhibition capacity of compounds to block NTCP substrate transportation,the equilibrium dissociation constant measured by SPR method can compare the binding affinity of compounds to NTCP,and can be used to develop anti-HBV drugs based on NTCP-PreS1 interaction.

关 键 词：钠离子/牛磺胆酸共转运多肽 乙肝病毒 表面等离子共振 间接偶联 动力学 

分 类 号：R917[医药卫生—药物分析学]