第5卷‧第3期,
198107
, pp. 273-277
臺灣產眼鏡蛇蛇毒磷脂?A2還原後?活性之氧化再生
- 作者:
陳長瑞; 吳世雄; 王光燦
- 作者服務機構:
中央研究院生化研究所
- 中文摘要:
臺灣產眼鏡蛇蛇毒磷脂?A2(Taiwan cobra phospholipase A2)在含有6M鹽酸脈的溶液中,經由2-硫酸乙醇的還
原作用將它所具有的七對雙硫鍵全部打斷,致使失去其原有的構造及?活性,當失去?活性的磷脂?在除去還原劑的情況
下可以慢慢再被氧化而恢復它原先的構造及?活性,在再氧化的過程中加入半胱氨酸不但可使恢復速度加快,也可使回收
量增加,加入半胱氨酸最佳的濃度是3mM,由再氧化回復後的磷脂?經過幾次的柱狀層析純化後,我們回收百分之六十的
磷脂?,它們不論是?活性,電泳性或CD光譜都和原來的磷脂?完全一致。
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- 英文摘要:
Reduction of Taiwan cobra (Naja naja atra) venom phospholipase A2 with 2-mercaptoethanol inthe presence of 6 M guanidine hydrochloride resulted in a complete disruption of all seven disulfide bridgesand loss of all enzymatic activity. The regeneration of enzymatic activity of fully reduced and inactivephospholipase A2 was found to be accelerated by cysteine and the optimal concentration was 3 mM. Afterpurification of the reoxidized phospholipase A2 by Sephadex G-50 gel filtration and CM-cellulose ionexchange chromatography, the pure phospholipase A2 was obtained in 60% yield. The purified regeneratedphospholipase A2 has the same specific activity, electromobility and circular dichroic spectrum as thoseof native phospholipase A2.
- 中文關鍵字:
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- 英文關鍵字:
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