具有抗白血病和淋巴瘤细胞增殖活性的菜豆胰蛋白酶抑制剂的分离与鉴定
Isolation and Characterization of a Phaseolus vulgaris Trypsin Inhibitor with Antiproliferative Activity on Leukemia and Lymphoma Cells.
作者信息
Li Miao, Liu Qin, Cui Yajuan, Li Dong, Wang Hexiang, Ng Tzi Bun
机构信息
State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing 100193, China.
Engineering Research Center of System-Nutrition, Beijing Research Institute for Nutritional Resources, Beijing 100069, China.
出版信息
Molecules. 2017 Jan 23;22(1):187. doi: 10.3390/molecules22010187.
A 17.5-kDa trypsin inhibitor was purified from cv. "gold bean" with an isolation protocol including ion exchange chromatography on DEAE-cellulose (Diethylaminoethyl-cellulose), affinity chromatography on Affi-gel blue gel, ion exchange chromatography on SP-sepharose (Sulfopropyl-sepharose), and gel filtration by FPLC (Fast protein liquid chromatography) on Superdex 75. It dose-dependently inhibited trypsin with an IC value of 0.4 μM, and this activity was reduced in the presence of dithiothreitol in a dose- and time-dependent manner, signifying the importance of the disulfide linkage to the activity. It inhibited [methyl-³H] thymidine incorporation by leukemia L1210 cells and lymphoma MBL2 cells with an IC value of 2.3 μM and 2.5 μM, respectively. The inhibitor had no effect on fungal growth and the activities of various viral enzymes when tested up to 100 μM.
从“金豆”品种中纯化出一种17.5 kDa的胰蛋白酶抑制剂，其分离方案包括在DEAE - 纤维素（二乙氨基乙基纤维素）上进行离子交换色谱、在Affi - 凝胶蓝胶上进行亲和色谱、在SP - 琼脂糖（磺丙基琼脂糖）上进行离子交换色谱以及通过Superdex 75上的快速蛋白质液相色谱（FPLC）进行凝胶过滤。它对胰蛋白酶的抑制作用呈剂量依赖性，IC值为0.4 μM，并且在二硫苏糖醇存在下，这种活性以剂量和时间依赖性方式降低，这表明二硫键对活性的重要性。它抑制白血病L1210细胞和淋巴瘤MBL2细胞掺入[甲基 - ³H]胸苷，IC值分别为2.3 μM和2.5 μM。当测试浓度高达100 μM时，该抑制剂对真菌生长和各种病毒酶的活性没有影响。
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