汞离子对木瓜蛋白酶结构的影响及抑制机理的研究

中国生物工程杂志

2012, Vol. 32

Issue (01): 87-91

研究简报

汞离子对木瓜蛋白酶结构的影响及抑制机理的研究

张艳梅, 曾虹燕, 蔡西玲, 熊龙斌, 张存滢, 侯茜

湘潭大学化工学院生物技术研究所湘潭 411105

Research of Hg²⁺ Effect on the Catalytic Activity of Papain and Its Inhibition Mechanism

ZHANG Yan-mei, ZENG Hong-yan, CAI Xi-ling, XIONG Long-bin, ZHANG Cun-ying, HOU Qian

Biotechnology Institute, College of Chemical Engineering, Xiangtan University, Xiangtan 411105, China

全文: PDF(653 KB) HTML

摘要：

通过荧光光谱、紫外光谱、傅里叶红外光谱(FTIR)和远紫外圆二色光谱(Far-UV CD)对重金属Hg²⁺作用下木瓜蛋白酶的结构变化进行了定性定量的表征,探索Hg²⁺对木瓜蛋白酶变性的抑制机理。结果表明:Hg²⁺是木瓜蛋白酶的强抑制剂,10^－4mol/L的HgCl₂可使木瓜蛋白酶丧失90%的活性;随着金属Hg²⁺浓度的增加,蛋白质所处的微环境和二级结构发生了明显的改变,(α螺旋＋β折叠)结构总量减少,无规则卷曲含量增多,酶的二级结构由规则有序转向无序。

关键词： 木瓜蛋白酶; Hg²⁺; 催化活性; 二级结构

Abstract:

Fluorescence spectroscopy, UV spectroscopy, Fourier transform infrared spectroscopy(FTIR) and Far UV circular dichroism (Far-UV CD) were done to characterize the structural changes of papain influenced by heavy metal ion Hg²⁺. And the denaturation and inhibition mechanisms of papain under the action of Hg²⁺were studied. The results indicated that Hg²⁺ was a strong inhibitor of papain. 90% enzymatic activity was lost at 10^－4mol/L Hg^{2 +}concentration. With Hg²⁺concentration increasing, the secondary structure and microenvironment of papain changed obviously. (α-helix + β-sheet) reduced, the content of random coil increased, and the secondary structure of the enzyme structure turned from regulation into disorder and random.

Key words: Papain Hg²⁺ Catalytic activity Secondary structure

收稿日期: 2011-09-06 出版日期: 2012-01-25

ZTFLH:

Q518.4

基金资助:

教育部大学生创新实验计划资助项目(1011053021)

	服务
	把本文推荐给朋友
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章

引用本文:

张艳梅, 曾虹燕, 蔡西玲, 熊龙斌, 张存滢, 侯茜. 汞离子对木瓜蛋白酶结构的影响及抑制机理的研究[J]. 中国生物工程杂志, 2012, 32(01): 87-91.

ZHANG Yan-mei, ZENG Hong-yan, CAI Xi-ling, XIONG Long-bin, ZHANG Cun-ying, HOU Qian. Research of Hg²⁺ Effect on the Catalytic Activity of Papain and Its Inhibition Mechanism. China Biotechnology, 2012, 32(01): 87-91.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2012/V32/I01/87

[1] 熊华.木瓜蛋白酶的应用研究进展. 保鲜与加工,2006 (1):7-8. Xiong H. The application and research dvelopment of papain.Storage & Process, 2006 (1):7-8.

[2] Kaul P, Sathish H A, Prakash V. Effect of metal ions on structure and activity of papain.. Nahrung/Food,2002,46(1):2-6.

[3] 何平,黄卓烈,初志战,等.乙腈对木瓜蛋白酶活性与构象的影响.上海交通大学学报,2006,24(3):245-249. He P,Huang Z L,Chu Z Z,et al.Effect of acetonitrile on activity and conformation of papain.Journal of ShangHai University, 2006,24(3):245-249.

[4] Jiang J,Yang X D,Wang K. Inhibition of cysteine protease papain by metal ions and polysulfide complexes, especially mercuric ion. Chinese Pharmaceutical Sciences,2007,16:1-8.

[5] Liu Y Y, Zhou H, Wang X Y, et al. Study on tryptophan modification and fluorescence spectrum of Sophora flavescens lectin. J Sichuan Univ Nat Sci Ed, 2001, 38(5):738- 742.

[6] 吴丹,徐桂英.光谱法研究蛋白质与表面活性剂的相互作用. 物理化学学报,2006,22 (2):254-260. Wu D,Xu G Y. Study on protein-surfactant interaction by spectroscopic methods.Acta Physico-Chimica Sinica, 2006,22 (2):254-260.

[7] Grzyska P K,Hausinger R P,Proshlyakov D A.Metal and substrate binding to an Fe(II) dioxygenase resolved by UV spectroscopy with global regression analysis.Analytical Biochemistry, 2010, 399(1):64-71.

[8] 叶宪曾,张新祥.仪器分析教程.北京:北京大学出版社,2007:24-27. Ye X Z,Zhang X X. Instrumental Analysis Tutorial. Beijing:Peking University Press,2007:24-27.

[9] Sebastian V, Jamie C, Heiko S. Implementation of an FTIR calibration curve for fast and objective determination of changes in protein secondary structure during formulation development. Pharmaceutical and Biomedical Analysis,2010,51:39-45.

[10] 沈琼, 黄滨, 邵嘉亮. 运用圆二色谱研究酶与化合物相互作用的机理.中山大学学报.2006,45(4):62-64. Shen Q, Huang B, Shao J L. Mechanism discussion of interaction between enzyme and several compounds with circular dichroism method.Journal of Sun Yat-sen University, 2006,45(4):62-64.

[11] Kelly S M,Jess T J,Price N C.How to study proteins by circular dichroism. Biochimica et Biophysica Acta, 2005, 1751:119-139.

[1]	阚婷婷,宗迅成,苏永君,王婷婷,李闯,胡蝶,邬敏辰. 定点突变改善PvEH1对邻甲基苯基缩水甘油醚的催化特性 ^*[J]. 中国生物工程杂志, 2019, 39(6): 9-16.
[2]	朱梦露,王雪雨,刘鑫,路福平,孙登岳,秦慧民. 一种新型亮氨酸5-羟化酶NmLEH的异源表达、纯化及酶学性质分析 ^*[J]. 中国生物工程杂志, 2019, 39(12): 24-34.
[3]	郭倩倩,高登科,程晓涛,路福平,田之仓优,秦慧民. 胆固醇氧化酶PsCO₄异源表达、纯化及酶学性质分析 ^*[J]. 中国生物工程杂志, 2018, 38(6): 34-42.
[4]	秦娇荣, 赵兆, 罗心梅, 李春阳. 基于mRNA 5'端TIR区二级结构优化提高重组sTNFα RI在大肠杆菌中的表达水平[J]. 中国生物工程杂志, 2018, 38(3): 62-69.
[5]	孙世琪,刘湘涛,郭慧琛,尹双辉,刘在新,马军武,谢庆阁. 猪水疱病病毒外壳蛋白的二级结构和抗原表位预测[J]. 中国生物工程杂志, 2006, 26(05): 1-6.
[6]	吕燕波, 万瑛, 吴玉章. SARS病毒M蛋白的二级结构和B细胞表位预测[J]. 中国生物工程杂志, 2003, 23(6): 41-45.
[7]	朱国萍, 滕脉坤, 王玉珍. 脯氨酸对蛋白质热稳定性的贡献[J]. 中国生物工程杂志, 2000, 20(4): 48-51.
[8]	刘新垣, 徐获, 蒋春雷, 王志勇, 郑仲承. 白细胞介素-2的结构与功能[J]. 中国生物工程杂志, 1994, 14(1): 13-17.
[9]	卓肇文. 抗体酶正在问世[J]. 中国生物工程杂志, 1992, 12(5): 46-49.
[10]	沈天翔, 贾盘兴. 质粒复制与反义RNA的调控机制[J]. 中国生物工程杂志, 1992, 12(2): 44-50.
[11]	马恩成. 用反义RNA和催化RNA抑制艾滋病毒[J]. 中国生物工程杂志, 1991, 11(2): 55-56.
[12]	卓肇文. 应用自然界的规律与化学家的工具是创造新蛋白质的一条途径[J]. 中国生物工程杂志, 1991, 11(1): 39-43.
[13]	杨俭华, 王露茜. 应用计算机预测核糖体16s rRNA三维折叠模型[J]. 中国生物工程杂志, 1990, 10(4): 18-21.
[14]	李祖义, 朱明华. 非水相生物催化[J]. 中国生物工程杂志, 1990, 10(2): 19-23.
[15]	TuckerJ.B., 刘桂生. 定做蛋白质[J]. 中国生物工程杂志, 1987, 7(3): 38-45.

Viewed

Full text

Abstract

Cited

Shared

Discussed