中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2008年
7期
582-586
,共5页
郑子峥%苗季%吴小成%何水珍%唐明%孙媛媛%王颖彬%杜海莲%张军%夏宁邵
鄭子崢%苗季%吳小成%何水珍%唐明%孫媛媛%王穎彬%杜海蓮%張軍%夏寧邵
정자쟁%묘계%오소성%하수진%당명%손원원%왕영빈%두해련%장군%하저소
戊型肝炎病毒%热休克蛋白90%蛋白相互作用
戊型肝炎病毒%熱休剋蛋白90%蛋白相互作用
무형간염병독%열휴극단백90%단백상호작용
Hepatitis E virus%HSP90%Protein-protein interaction
目的 对前期研究中所筛选出的能够与戊型肝炎病毒(hepatitis E virus,HEV)重组衣壳蛋白颗粒P239存在特异性的相互作用的蛋白进行分析验证.方法 使用pull-down、MALDI-TOF-MS、免疫共沉淀技术及免疫荧光技术对与P239有潜在相互作用的蛋白进行鉴定和进一步分析.结果 MALDI-TOF-MS及免疫共沉淀技术,均表明该蛋白为热休克蛋白90(HSP90)并与P239相互作用.随后,对P239进入细胞的过程中胞内HSP90蛋白量及其分布的变化做了初步研究.发现虽然HSP90在蛋白总量上并没有发生明显的变化,但其定位却伴随着P239在胞内的迁移发生了由细胞膜向细胞核核周的转移.结论 HSP90可能参与了P239入胞后的转运并介导了HEV入胞后向效应细胞器的转运,为研究HEV的感染机制及对HEV的预防和控制提供了有益的线索.
目的 對前期研究中所篩選齣的能夠與戊型肝炎病毒(hepatitis E virus,HEV)重組衣殼蛋白顆粒P239存在特異性的相互作用的蛋白進行分析驗證.方法 使用pull-down、MALDI-TOF-MS、免疫共沉澱技術及免疫熒光技術對與P239有潛在相互作用的蛋白進行鑒定和進一步分析.結果 MALDI-TOF-MS及免疫共沉澱技術,均錶明該蛋白為熱休剋蛋白90(HSP90)併與P239相互作用.隨後,對P239進入細胞的過程中胞內HSP90蛋白量及其分佈的變化做瞭初步研究.髮現雖然HSP90在蛋白總量上併沒有髮生明顯的變化,但其定位卻伴隨著P239在胞內的遷移髮生瞭由細胞膜嚮細胞覈覈週的轉移.結論 HSP90可能參與瞭P239入胞後的轉運併介導瞭HEV入胞後嚮效應細胞器的轉運,為研究HEV的感染機製及對HEV的預防和控製提供瞭有益的線索.
목적 대전기연구중소사선출적능구여무형간염병독(hepatitis E virus,HEV)중조의각단백과립P239존재특이성적상호작용적단백진행분석험증.방법 사용pull-down、MALDI-TOF-MS、면역공침정기술급면역형광기술대여P239유잠재상호작용적단백진행감정화진일보분석.결과 MALDI-TOF-MS급면역공침정기술,균표명해단백위열휴극단백90(HSP90)병여P239상호작용.수후,대P239진입세포적과정중포내HSP90단백량급기분포적변화주료초보연구.발현수연HSP90재단백총량상병몰유발생명현적변화,단기정위각반수착P239재포내적천이발생료유세포막향세포핵핵주적전이.결론 HSP90가능삼여료P239입포후적전운병개도료HEV입포후향효응세포기적전운,위연구HEV적감염궤제급대HEV적예방화공제제공료유익적선색.
Objective To identify the protein interacting with hepatitis E virus(HEV) recombi-nant capsomeric particles(P239). Methods Protein interacting with HEV was analyzed by the pull-down, MALDI-TOF-MS, co-immunoprecipitation (Co-IP) and CONFOCAL. Results A protein interacting with HEV recombinant particle (P239) was identified as HSP90 by MALDI-TOF-MS. The interaction between HSP90 and P239 was further confirmed by Co-IP. The protein level and localization of HSP90 and P239 in HepG2 were detected. The total quantity of HSP90 didn't change, and the movement of HSP90 from plasma membrane to perinuclei region with P239 was observed. Conclusion HSP90 may play an important role in the trafficking of P239. It suggests that HSP90 participate in the transportation of HEV after infection, which may contribute to the prevention and control of the disease.
