【摘要】 明确SURFIN_4.1蛋白氨基（N）端在转运过程中的作用。通过恶性疟原虫转染筛选表达SURFIN_4.1^2Myc-N-T-Cyt重组蛋白的MS822转染株;通过IFA和Western Blot方法对比分析N端Myc标签对SURFIN_4.1重组蛋白定位和可溶性影响;通过免疫共沉淀（Co-IP）和质谱分析（LC-MS/MS）方法,明确SURFIN_4.1蛋白N端转运过程中的相互作用蛋白。成功筛选并获得表达SURFIN_4.1^2Myc-N-T-Cyt重组蛋白的恶性疟原虫MS822转染株;SURFIN_4.1^2Myc-N-T-Cyt与SURFIN_4.1^N-T-Cyt的定位和可溶性无明显差异。Co-IP和LC-MS/MS结果显示,SURFIN_4.1蛋白N端与茂氏点定位蛋白MAHRP1,PTEX复合体成员EXP2,棒状体蛋白RAP3,红细胞膜表面多种原虫蛋白及HSP60相互作用。结果表明,SURFIN_4.1蛋白N端在转运过程中不水解,且N端Myc标签不影响蛋白可溶性和定位;SURFIN_4.1蛋白N端与多种恶性疟致病性相关蛋白相互作用,提示SURFIN_4.1蛋白作为红内期疫苗候选抗原研制靶位的可能性。更多还原

【Abstract】 The role of SURFIN_4.1N-terminal during protein transportation was investigated. The MS822 transgenic parasites that express recombinant SURFINSURFIN_4.1^2Myc-N-T-Cytwas generated by P. falciparum transfection. Indirect immunofluorescence and western blot assays were performed to compare and analyze the N-terminal tagging influence on location and solubility. The N-terminal interaction proteins were detected by Co-IP and LC-MS / MS. The MS822 transgenic parasites that express recombinant SURFINSURFIN_4.1^2Myc-N-T-Cytwere successfully generated. No significant location and solubility variations were detected after N-terminal tagging of SURFIN_4.1. Co-IP and LC-MS / MS detected Maurer’s cleft protein MAHRP1,PTEX components EXP2,rhabdoid protein RAP3,pRBCs surface proteins,and HSP60 interact with N-terminal of SURFIN_4.1. The N-terminal of SURFIN_4.1did not hydrolyze during transportation,and N-terminal Myc tagging did not affect protein location and solubility. And N-terminal of SURFIN_4.1interacted with several parasite pathogenicity related proteins prompted that the possibility of SURFIN_4.1protein to develop target location for vaccine candidate antigen in erythrocytic stage.更多还原