| 摘要: |
| 以重组人tPA蛋白为材料研究了精氨酸、精氨酸盐酸盐、半胱氨酸、胱氨酸对蛋白质复性效果的影响,重组tPA蛋白包涵体经尿素变性溶解后,在精氨酸、精氨酸盐酸盐、半胱氨酸、胱氨酸存在的条件下进行复性,结果表明,碱性的精氨酸在质量分数0.2%时可减少蛋白质凝聚,显著提高复性效果,tPA复性后的活性可提高50%以上,半胱氨酸单独使用具有类似β-巯基乙醇的作用,精氨酸盐酸盐和胱氨酸单独使用对复性无影响,而半胱氨酸和胱氨酸联合使用,有类似氧化-还原系统作用.可提高活性20%. |
| 关键词: 重组tPA 复性 |
| DOI： |
| 分类号:Q503 |
| 基金项目:国家科技攻关计划，湖北省科研项目 |
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| Effects of Arginine, Arginine Hydrochloride, Cysteine, Cystine on tPA Renaturation |
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| Abstract: |
| The effects of arginine,arginine-cl,cysteine and cystine on the renaturation of human tPA were studied.tPA inclusion body protein was dissolved and denatured in 8M urea with 5mM 2-ME at pH 9.5.After removed urea and 2-ME by G-25 column chromatography,the tPA protein in the solution was renatured in the present of arginine,arginine-cl,cysteine and cysteine-cystine.At pH 8.0 and room temperature for 24hr,the efficiency of renaturation was evaluated according to the activity of tPA.The results demonstrated that the renaturation efficiency of tPA was increased over 50﹪by arginine alone,but arginine hydrochloride and cystine had no effect on tPA renaturation,the cysteine alone or cysteine plus cystine could slightly increase tPA activity. |
| Key words: recombinant tPA renaturation |
