Department of Biotechnology, School of Bioengineering, SRM University, Kattankulathur 603203, India.
Biomed Res Int. 2014;2014:179836. doi: 10.1155/2014/179836. Epub 2014 Jan 30.
A 35 KDa protein referred to as F3 was purified from the seeds of Borreria hispida by precipitation with 80% ammonium sulphate and gel filtration on Sephadex G-100 column. RP-HPLC analysis of protein fraction (F3) on an analytical C-18 column produced a single peak, detected at 220 nm. F3 showed an apparent molecular weight of 35 KDa by SDS PAGE and MALDI-TOF-MS analyses. Peptide mass fingerprinting analysis of F3 showed the closest homology with the sequence of 1-aminocyclopropane-1-carboxylate deaminase of Pyrococcus horikoshii. The protein (F3) exhibited significant cytotoxic activity against lung (A549) and cervical (HeLa) cancer cells in a dose-dependent manner at concentrations ranging from 10 µg to 1000 µg/mL, as revealed by the MTT assay. Cell cycle analysis revealed the increased growth of sub-G0 population in both cell lines exposed to a concentration of 1000 µg/mL of protein fraction F3 as examined from flow cytometry. This is the first report of a protein from the seeds of Borreria hispida with antiproliferative and apoptotic activity in lung (A549) and cervical (HeLa) cancer cells.
一种 35 kDa 的蛋白质 F3 被从 Borreria hispida 的种子中提取出来,方法是用 80%的硫酸铵沉淀和 Sephadex G-100 柱凝胶过滤。对蛋白质部分(F3)在分析性 C-18 柱上进行 RP-HPLC 分析,产生一个单一的峰,在 220nm 处检测到。SDS PAGE 和 MALDI-TOF-MS 分析表明,F3 的表观分子量为 35 kDa。F3 的肽质量指纹图谱分析显示与 Pyrococcus horikoshii 的 1-氨基环丙烷-1-羧酸脱氨酶序列具有最接近的同源性。该蛋白(F3)在浓度为 10μg 至 1000μg/mL 时,对肺(A549)和宫颈(HeLa)癌细胞表现出显著的细胞毒性活性,这一点通过 MTT 测定得到证实。细胞周期分析显示,在暴露于 1000μg/mL 蛋白部分 F3 的两种细胞系中,亚 G0 群体的生长增加,这是从流式细胞术检查中发现的。这是首次报道来自 Borreria hispida 种子的一种具有抗增殖和促凋亡活性的蛋白质,可作用于肺(A549)和宫颈(HeLa)癌细胞。