Song Liyan, Ren Shengfang, Yu Rongmin, Yan Chunyan, Li Tingfei, Zhao Yu
College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, P.R. China.
Mar Drugs. 2008;6(3):418-30. doi: 10.3390/md20080020. Epub 2008 Jul 10.
Two purified proteins G-6 and G-4-2 were obtained from Arca subcrenata Lischke using the homogenization, salting-out with ammonium sulfate, ion-exchange chromatography and gel filtration chromatography techniques. The purity of G-6 and G-4-2 was over 96%, as measured by RP-HPLC. G-6 and G-4-2 were measured by SDS-PAGE and IEF-PAGE to have molecular weights of 8.2 kDa and 16.0 kDa, and isoelectric points of 6.6 and 6.1, respectively. The amino acid constituents of G-6 and G-4-2 were also determined. The existence of saccharides in G-6 was demonstrated by the phenol-sulfuric acid method. G-6 and G-4-2 inhibited the proliferation of human tumor cells in vitro. By MTT assay, the IC(50) values of G-4-2 were 22.9 microg/mL, 46.1 microg/mL and 57.7 microg/mL against Hela, HL-60 and KB cell lines, respectively, and the IC(50) value of G-6 against HL-60 cell line was measured to be 123.2 microg/mL.
采用匀浆、硫酸铵盐析、离子交换色谱和凝胶过滤色谱技术，从皱纹盘鲍中获得了两种纯化蛋白G-6和G-4-2。通过反相高效液相色谱（RP-HPLC）测定，G-6和G-4-2的纯度超过96%。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和等电聚焦电泳（IEF-PAGE）测定，G-6和G-4-2的分子量分别为8.2 kDa和16.0 kDa，等电点分别为6.6和6.1。还测定了G-6和G-4-2的氨基酸组成。通过苯酚-硫酸法证明了G-6中存在糖类。G-6和G-4-2在体外抑制人肿瘤细胞的增殖。通过MTT法测定，G-4-2对Hela、HL-60和KB细胞系的半数抑制浓度（IC50）值分别为22.9 μg/mL、46.1 μg/mL和57.7 μg/mL，G-6对HL-60细胞系的IC50值测定为123.2 μg/mL。
