蓝藻丝氨酸蛋白酶抑制剂arthropin的制备及其靶蛋白酶的筛选

许佳伟, 周爱武, 杨愈丰

Preparation and target protease identification of a cyanobacterial serine protease inhibitor, arthropin

XU Jiawei, ZHOU Aiwu, YANG Yufeng

图2 SDS-PAGE分析arthropin融合蛋白的表达、酶切与纯化 Note：A. Protein expression in the whole bacteria and lysis supernatant of E. coli before and after induction. Lane 1—marker; lane 2—total bacterial protein sample before induction; lane 3—supernatant protein sample before induction; lane 4—total bacterial protein sample after induction for 12 h; lane 5—supernatant protein sample after induction for 12 h. B. Enzymatic digestion and purification by nickel ion affinity chromatography. Lane 1—marker; lane 2—protein sample before SENP2 digestion; lane 3—protein sample after SENP2 digestion; lane 4?5—flow-through solution of the nickel column after SENP2 digestion; lane 6?7—washing solution of the nickel column after SENP2 digestion; lane 8—elution solution of the nickel column after SENP2 digestion. C. Purification by anion-exchange chromatography. Lane 1—marker; lane 2?12—protein samples eluted by a continuous gradient of 145?685 mmol/L NaCl. The black arrow indicates the fusion protein and the red arrows indicate the monomeric protein.

Fig 2 Expression, enzymatic digestion and purification of arthropin fusion protein analyzed by SDS-PAGE