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。
This report described the purification of a 30 kDa ribonuclease （RNase） from dry fruiting bodies of the mushroom Russu- la aeruginea Lindb.： Fr. The following chromatographic steps were employed sequentially： ion exchange chromatography on CM-cel- lulose, 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 ac- tivity （13.7U·mg^-1） toward poly C, and much weaker activity toward poly G （1.2U·mg^-1） and poly U （0.7 U·mg^-1） . The RNase was significantly inhibited by Hg^2＋ , Pb^2＋ , Fe^2＋ and Al^3＋ ions. It inhibited HIV-1 reverse transcriptase and proliferation of hepatoma Hep G2 cells with an ICso 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 1.4μmol·L^-1, signifying that it was an antipathogenic protein.
出处
《中国食用菌》
2013年第2期28-32,共5页
Edible Fungi of China
基金
北京市博士后科研活动工作经费(2012ZZ-33)
关键词
核糖核酸酶
铜绿红菇
HIV-1
抗增殖
Ribonuclease
Russula aeruginea Lindb.： Fr.
HIV-1
Antiproliferation
