State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing 100193, P.R. China.
Int J Mol Med. 2014 Jan;33(1):209-14. doi: 10.3892/ijmm.2013.1553. Epub 2013 Nov 12.
In this study, a 27-kDa ribonuclease (RNase) was purified from the dried fruiting bodies of the mushroom, Hohenbuehelia serotina. The isolation protocol involved anion exchange chromatography, affinity chromatography, cation exchange chromatography and gel filtration in succession. The RNase was unadsorbed on DEAE-cellulose, but was adsorbed on Affi-gel blue gel and CM-cellulose. The N-terminal amino acid sequence was TVGGSLAEKGN which showed homology to other fungal RNases to a certain degree. The RNase exhibited maximal RNase activity at pH 5 and 80˚C. It demonstrated the highest ribonucleolytic activity toward poly(C), a relatively high activity toward poly(U), and a considerably weaker activity toward poly(A) and (G). The RNase inhibited human immunodeficiency virus type 1 (HIV-1) reverse transcriptase with an IC50 of 50 µM and reduced [3H-methyl]-thymidine uptake by L1210 leukemia cells and MBL2 lymphoma cells with an IC50 of 25 µM and 40 µM, respectively.
本研究从白蘑科真菌 Hohenbuehelia serotina 的干燥子实体中纯化得到一种 27kDa 的核糖核酸酶（RNase）。该分离方案依次包括阴离子交换层析、亲和层析、阳离子交换层析和凝胶过滤。RNase 不被 DEAE-纤维素吸附，但被 Affi-gel blue 凝胶和 CM-纤维素吸附。RNase 的 N 端氨基酸序列为 TVGGSLAEKGN，与其他真菌 RNase 具有一定程度的同源性。RNase 在 pH5 和 80°C 时表现出最大的 RNase 活性。它对聚（C）表现出最高的核糖核酸酶活性，对聚（U）具有相对较高的活性，对聚（A）和（G）的活性则较弱。RNase 以 50µM 的 IC50 抑制人类免疫缺陷病毒 1（HIV-1）逆转录酶，并以 25µM 和 40µM 的 IC50 分别降低 L1210 白血病细胞和 MBL2 淋巴瘤细胞中[3H-甲基]胸苷的摄取。
