CAJ | 학술논문

目的明确4-1 BB分子的配体结合4-1 BB分子胞外区的关键区域和基本结构.方法分区表达鼠4-1 BB胞外区结构域及其天然配体,利用ELISA和Western blot等方法对4-1 BB分子的配体识别结构域进行定位.结果 4-1 BB分子半胱氨酸寓集结构域(cysteine-rich domain,CRD)Ⅱ是配体结合的主要结构域,不结合CRD Ⅰ和CRDⅢ.免疫刺激性抗4-1 BB单克隆抗体2A,主要结合4-1 BB分子CRD与跨膜区之间的连接区(STP区),同时对包括CRDⅡ的各个CRDs有弱识别,可封阻4-1 BB与其配体结合.结论 4-1 BB分子配体的主要识别区位于4-1 BB分子胞外CRD Ⅱ,具有空间结构特点,此为进一步分析结合区及其关键氨基酸提供了资料.
목적 명학4-1 BB분자적배체결합4-1 BB분자포외구적관건구역화기본결구.방법 분구표체서4-1 BB포외구결구역급기천연배체,이용ELISA화Western blot등방법 대4-1 BB분자적배체식별결구역진행정위.결과 4-1 BB분자반광안산우집결구역(cysteine-rich domain,CRD)Ⅱ시배체결합적주요결구역,불결합CRD Ⅰ화CRDⅢ.면역자격성항4-1 BB단극륭항체2A,주요결합4-1 BB분자CRD여과막구지간적련접구(STP구),동시대포괄CRDⅡ적각개CRDs유약식별,가봉조4-1 BB여기배체결합.결론 4-1 BB분자배체적주요식별구위우4-1 BB분자포외CRD Ⅱ,구유공간결구특점,차위진일보분석결합구급기관건안기산제공료자료.
ObJective To locate the cysteine-rich domains(CRD) of murine 4-1BB binding to its natural ligand. Methods A serial soluble extracellular CRDs of routine 4-1BB and 4-1BBL fusion proteins was constructed and prepared. The binding of purified 4-1BB-Igs to 4-1BBL and 4-1BB monoclonal antibody were tested using ELISA assay and Western blot analysis. Blocking experiment with 4-1BBL and 4-1BB mon-oclonal antibody was performed by ELISA assay. Results All truncated overlapped proteins containing ex-tracellular CRD Ⅱ of murine 4-1BB were able to bind to 4-1BBL by ELISA assay, excepting the CRD Ⅰ do-main alone. A 4-1BB monoclonal antibody proved to block the interaction of 4-1BB and 4-1BBI, was also able to bind to CRD Ⅱ. Conclusion Murine 4-1BBL whose specificity was mapped to CRD Ⅱ of 4-1BB ex-tracellular region with a possible conformational structure.