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一种具有增殖活性的铜绿红菇核糖核酸酶

作  者:
李淼;李东;王贺祥
单  位:
中国农业大学生物学院农业生物技术国家重点实验室;北京市营养源研究所分析室
关键词:
核糖核酸酶;铜绿红菇;HIV-1;抗增殖
摘  要:
本研究采用CM-cellulose、Q-sepharose离子交换层析、Superdex 75凝胶过滤层析等蛋白质分离纯化方法,以铜绿红菇的干子实体为材料,分离纯化出一种新的分子量为30 kDa的核糖核酸酶,该酶最适反应温度为60℃,最适反应pH为5左右。该酶对寡聚腺苷酸的降解活性最强,为15.4 U.mg-1,对寡聚胞嘧啶的降解活性次之,为13.7 U.mg-1,对寡聚鸟嘌呤和寡聚尿嘧啶的降解活性最弱,分别为1.2 U.mg-1和0.7 U.mg-1。大部分金属离子对该酶的活性有抑制作用,以Hg2+、Pb2+、Fe2+和Al3+对其抑制作用最为明显。该酶能够抑制HIV-1逆转录酶活性,IC50为1μmol.L-1,体外抑制人肝癌细胞株Hep G2和小鼠白血病细胞株L1210细胞系增殖的IC50分别为3.1μmol.L-1和1.4μmol.L-1。
译  名:
A Novel Ribonuclease with Antiproliferative Activity from Russula aeruginea Lindb.:Fr.
作  者:
LI Miao1,2,LI Dong1,WANG He-xiang2 (1.Analysis Laboratory of Beijing Research Institute for Nutritional Resources,Beijing 100069;2.State Key Laboratory for Agrobiotechnology and Department of Microbiology,China Agricultural University,Beijing 100193)
关键词:
Ribonuclease;Russula aeruginea Lindb.:Fr.;HIV-1;Antiproliferation
摘  要:
This report described the purification of a 30kDa ribonuclease(RNase) from dry fruiting bodies of the mushroom Russula aeruginea Lindb.:Fr..The following chromatographic steps were employed sequentially: ion exchange chromatography on CM-cellulose,Q-sepharose,and gel filtration by fast protein liquid chromatography on Superdex 75.The RNase exhibited maximal RNase activity at pH 5 and 60℃.It demonstrated the highest ribonucleolytic activity(15.4 U·mg-1) toward poly A,the next highest activity(13.7 U·mg-1) toward poly C,and much weaker activity toward poly G(1.2 U·mg-1) and poly U(0.7 U·mg-1).The RNase was significantly inhibited by Hg2+,Pb2+,Fe2+and Al3+ ions.It inhibited HIV-1 reverse transcriptase and proliferation of hepatoma Hep G2 cells with an IC50 of 1 μmol·L-1 and 3.1 μmol·L-1,respectively.The RNase curtailed [3-Hmethyl]-thymidine uptake by leukemia L1210 cells with an IC50 of 1.4 μmol·L-1,signifying that it was an antipathogenic protein.

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