静电排斥表面诱导溶菌酶分子站立

引用本文: 白姝, 李浩, 张麟. 静电排斥表面诱导溶菌酶分子站立[J]. 物理化学学报, doi: 10.3866/PKU.WHXB201301182 shu

Citation: BAI Shu, LI Hao, ZHANG Lin. Standing Orientation of Lysozymes Induced by Electrostatically Repulsive Surface[J]. Acta Physico-Chimica Sinica, doi: 10.3866/PKU.WHXB201301182 shu

静电排斥表面诱导溶菌酶分子站立

基金项目:
国家自然科学基金(21236005, 21006069) (21236005, 21006069)

天津大学自主创新基金资助项目

摘要:

抑制蛋白质聚集是应用基因重组技术生产药用蛋白质过程中的关键. 实验研究发现与蛋白质带同种电荷的离子交换介质能够通过静电排斥作用有效抑制蛋白质折叠中间体的聚集. 但其微观细节尚不明晰, 且利用现有实验技术很难直接阐释. 分子动力学模拟是研究微观过程的有力工具. 因此, 本文构建了静电排斥表面模型以模拟同电荷离子交换介质, 采用分子动力学模拟和全原子模型, 研究溶菌酶在静电排斥表面上的空间取向及其变化过程, 并考察表面所带电荷数的影响规律. 结果表明, 溶菌酶受到表面的静电排斥作用而远离. 在此过程中, 溶菌酶逐渐“站立”, 形成其偶极和表面相站立垂直的空间取向. 而当蛋白质远离表面时, 由于静电排斥作用衰减, 形成“站立”取向的趋势减弱. 同时, 研究发现静电排斥表面所带电荷数增加有利于蛋白质形成“站立”取向. 本文的模拟结果从微观揭示蛋白质在静电排斥表面上的空间取向及其影响因素, 将有助于推动蛋白质在荷电表面折叠和分子相互作用研究.

关键词:

English

Standing Orientation of Lysozymes Induced by Electrostatically Repulsive Surface

1.
1 Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, P. R. China;
2 Key Laboratory of Systems Bioengineering, Ministry of Education, Tianjin University, Tianjin 300072, P. R. China
Received Date: 22 October 2012
Available Online: 25 March 2013
Fund Project:
国家自然科学基金(21236005, 21006069)

天津大学自主创新基金资助项目

Abstract:

Inhibition of protein aggregation during protein refolding is a fundamental issue in the production of recombinant therapeutic proteins. It has recently been experimentally found that like-charged ion-exchange resin can suppress the aggregation of folding intermediates through electrostatic repulsion. However, the microscopic understanding of this process is far from adequate, and it is difficult to examine the microscopic process using experimental approaches. Molecular dynamics (MD) simulation is a powerful tool which can provide clear microscopic information in a direct manner. Therefore, in the present study, a model of an electrostatically repulsive surface is constructed to simulate the like-charged ion-exchange resin. The distribution of lysozyme molecules near the surface is then investigated using MD simulation with all-atom (AA) models and the effect of the charge number of the surface is examined. It is found that the protein is excluded from the surface by electrostatic repulsion. During this process, the protein molecule becomes standing, where the dipole of the protein is perpendicular to the surface. When the protein moves far from the surface, diminished oriented alignment is observed due to the decreased electrostatic repulsion from the surface. It is also found that better oriented alignment on the surface occurs with higher charge number. The simulation results thus provide microscopic information about the alignment of protein molecules near an electrostatically repulsive surface, and are expected to be helpful for investigation of protein refolding on charged surfaces and the molecular interactions involved.

Key words:

1. [1]
  
  (1) Durocher, Y.; Butler, M. Curr. Opin. Biotech. 2009, 20, 700. doi: 10.1016/j.copbio.2009.10.008
  (1) Durocher, Y.; Butler, M. Curr. Opin. Biotech. 2009, 20, 700. doi: 10.1016/j.copbio.2009.10.008
2. [2]
  (2) Manning, M. C.; Chou, D. K.; Murphy, B. M.; Payne, R.W.;Katayama, D. S. Pharm. Res. 2010, 27, 544. doi: 10.1007/s11095-009-0045-6(2) Manning, M. C.; Chou, D. K.; Murphy, B. M.; Payne, R.W.;Katayama, D. S. Pharm. Res. 2010, 27, 544. doi: 10.1007/s11095-009-0045-6
3. [3]
  (3) Ferrer-Miralles, N.; Domin -Espin, J.; Corchero, J. L.;Vazquez, E.; Villaverde, A. Microb. Cell. Fact. 2009, 8, 17. doi: 10.1186/1475-2859-8-17(3) Ferrer-Miralles, N.; Domin -Espin, J.; Corchero, J. L.;Vazquez, E.; Villaverde, A. Microb. Cell. Fact. 2009, 8, 17. doi: 10.1186/1475-2859-8-17
4. [4]
  (4) Fink, A. L. Fold. Des. 1998, 3, R9.(4) Fink, A. L. Fold. Des. 1998, 3, R9.
5. [5]
  (5) Middelberg, A. R. Trends Biotechnol. 2002, 20, 437. doi: 10.1016/S0167-7799(02)02047-4(5) Middelberg, A. R. Trends Biotechnol. 2002, 20, 437. doi: 10.1016/S0167-7799(02)02047-4
6. [6]
  (6) Baldwin, R. L. J. Mol. Biol. 2007, 371, 283. doi: 10.1016/j.jmb.2007.05.078(6) Baldwin, R. L. J. Mol. Biol. 2007, 371, 283. doi: 10.1016/j.jmb.2007.05.078
7. [7]
  (7) Chen, Y.W.; Ding, F.; Nie, H. F.; Serohijos, A.W.; Sharma, S.;Wilcox, K. C.; Yin, S. Y.; Dokholyan, N. V. Arch. Biochem.Biophys. 2008, 469, 4. doi: 10.1016/j.abb.2007.05.014(7) Chen, Y.W.; Ding, F.; Nie, H. F.; Serohijos, A.W.; Sharma, S.;Wilcox, K. C.; Yin, S. Y.; Dokholyan, N. V. Arch. Biochem.Biophys. 2008, 469, 4. doi: 10.1016/j.abb.2007.05.014
8. [8]
  (8) Lu, D.; Liu, Z. Annu. Rep. Prog. Chem., Sect. C 2010, 106, 259.doi: 10.1039/b903487k(8) Lu, D.; Liu, Z. Annu. Rep. Prog. Chem., Sect. C 2010, 106, 259.doi: 10.1039/b903487k
9. [9]
  (9) Wang, G.; Dong, X.; Sun, Y. Biotechnol. Bioeng. 2011, 108,1068. doi: 10.1002/bit.23038(9) Wang, G.; Dong, X.; Sun, Y. Biotechnol. Bioeng. 2011, 108,1068. doi: 10.1002/bit.23038
10. [10]
  (10) Karplus, M. Biopolymers 2003, 68, 350.(10) Karplus, M. Biopolymers 2003, 68, 350.
11. [11]
  (11) Karplus, M.; McCammon, J. A. Nat. Struct. Biol. 2002, 9, 646.doi: 10.1038/nsb0902-646(11) Karplus, M.; McCammon, J. A. Nat. Struct. Biol. 2002, 9, 646.doi: 10.1038/nsb0902-646
12. [12]
  (12) Bai, H. J.; Lai, L. H. Acta Phys. -Chim. Sin. 2010, 26, 1988.[白红军, 来鲁华. 物理化学学报, 2010, 26, 1988.] doi: 10.3866/PKU.WHXB20100725(12) Bai, H. J.; Lai, L. H. Acta Phys. -Chim. Sin. 2010, 26, 1988.[白红军, 来鲁华. 物理化学学报, 2010, 26, 1988.] doi: 10.3866/PKU.WHXB20100725
13. [13]
  (13) Zhang, X. R.;Wang,W. C. Acta Phys. -Chim. Sin. 2002, 18,680. [张现仁, 汪文川. 物理化学学报, 2002, 18, 680.]doi: 10.3866/PKU.WHXB20020803(13) Zhang, X. R.;Wang,W. C. Acta Phys. -Chim. Sin. 2002, 18,680. [张现仁, 汪文川. 物理化学学报, 2002, 18, 680.]doi: 10.3866/PKU.WHXB20020803
14. [14]
  (14) Xiang, Z. H.;Wang,W. C.; Cao, D. P. Scientia Sinica Chimica2012, 42, 235. [向中华, 汪文川, 曹达鹏. 中国科学: 化学,2012, 42, 235.] doi: 10.1360/032011-297(14) Xiang, Z. H.;Wang,W. C.; Cao, D. P. Scientia Sinica Chimica2012, 42, 235. [向中华, 汪文川, 曹达鹏. 中国科学: 化学,2012, 42, 235.] doi: 10.1360/032011-297
15. [15]
  (15) Kubiak-Ossowska, K.; Mulheran, P. A. Langmuir 2010, 26,15954. doi: 10.1021/la102960m(15) Kubiak-Ossowska, K.; Mulheran, P. A. Langmuir 2010, 26,15954. doi: 10.1021/la102960m
16. [16]
  (16) Ravichandran, S.; Madura, J. D.; Talbot, J. J. Phys. Chem. B2001, 105, 3610. doi: 10.1021/jp010223r(16) Ravichandran, S.; Madura, J. D.; Talbot, J. J. Phys. Chem. B2001, 105, 3610. doi: 10.1021/jp010223r
17. [17]
  (17) Carlsson, F.; Hyltner, E.; Arnebrant, T.; Malmsten, M.; Linse, P.J. Phys. Chem. B 2004, 108, 9871. doi: 10.1021/jp0495186(17) Carlsson, F.; Hyltner, E.; Arnebrant, T.; Malmsten, M.; Linse, P.J. Phys. Chem. B 2004, 108, 9871. doi: 10.1021/jp0495186
18. [18]
  (18) Slater, G.W.; Holm, C.; Chubynsky, M. V.; de Haan, H. H.;Dube, A.; Grass, K.; Hickey, O. A.; Kingsburry, C.; Sean, D.;Shendruk, T. N.; Nhan, L. X. Electrophoresis 2009, 30, 792.doi: 10.1002/elps.v30:5(18) Slater, G.W.; Holm, C.; Chubynsky, M. V.; de Haan, H. H.;Dube, A.; Grass, K.; Hickey, O. A.; Kingsburry, C.; Sean, D.;Shendruk, T. N.; Nhan, L. X. Electrophoresis 2009, 30, 792.doi: 10.1002/elps.v30:5
19. [19]
  (19) Chandrasekhar, I.; Kastenholz, M.; Lins, R. D.; Oostenbrink, C.;Schuler, L. D.; Tieleman, D. P.; van Gunsteren,W. F. Eur.Biophys. J. Biophy. 2003, 32, 67.(19) Chandrasekhar, I.; Kastenholz, M.; Lins, R. D.; Oostenbrink, C.;Schuler, L. D.; Tieleman, D. P.; van Gunsteren,W. F. Eur.Biophys. J. Biophy. 2003, 32, 67.
20. [20]
  (20) Zhang, L.; Zhao, G. F.; Sun, Y. J. Phys. Chem. B 2009, 113,6873. doi: 10.1021/jp809754k(20) Zhang, L.; Zhao, G. F.; Sun, Y. J. Phys. Chem. B 2009, 113,6873. doi: 10.1021/jp809754k
21. [21]
  (21) Zhang, L.; Zhao, G. F.; Sun, Y. J. Phys. Chem. B 2010, 114,2203. doi: 10.1021/jp903852c(21) Zhang, L.; Zhao, G. F.; Sun, Y. J. Phys. Chem. B 2010, 114,2203. doi: 10.1021/jp903852c
22. [22]
  (22) Kang, K.; Lu, D. N.; Liu, Z. Chin. J. Chem. Eng. 2012, 20, 284.(22) Kang, K.; Lu, D. N.; Liu, Z. Chin. J. Chem. Eng. 2012, 20, 284.
23. [23]
  (23) Shen, L. M.; Lin, D. Q.; Mei, L. H.; Yao, S. J. Chinese Journalof Bioprocess Engineering 2006, 4, 29. [沈立民, 林东强,梅乐和, 姚善泾. 生物加工过程, 2006, 4, 29.](23) Shen, L. M.; Lin, D. Q.; Mei, L. H.; Yao, S. J. Chinese Journalof Bioprocess Engineering 2006, 4, 29. [沈立民, 林东强,梅乐和, 姚善泾. 生物加工过程, 2006, 4, 29.]
24. [24]
  (24) Diamond, R. J. Mol. Biol. 1974, 82, 371. doi: 10.1016/0022-2836(74)90598-1(24) Diamond, R. J. Mol. Biol. 1974, 82, 371. doi: 10.1016/0022-2836(74)90598-1
25. [25]
  (25) Schuttelkopf, A.W.; van Aaltern, D. M. F. Acta Crystallogr. D2004, 60, 1355. doi: 10.1107/S0907444904011679(25) Schuttelkopf, A.W.; van Aaltern, D. M. F. Acta Crystallogr. D2004, 60, 1355. doi: 10.1107/S0907444904011679
26. [26]
  (26) Berendsen, H. J.; van der Spoel, D.; van Drunen, R. Comput. Phys.Commun. 1995, 91, 43. doi: 10.1016/0010-4655(95)00042-E(26) Berendsen, H. J.; van der Spoel, D.; van Drunen, R. Comput. Phys.Commun. 1995, 91, 43. doi: 10.1016/0010-4655(95)00042-E
27. [27]
  (27) Lindahl, E.; Hess, B.; van der Spoel, D. J. Mol. Model. 2001, 7,306.(27) Lindahl, E.; Hess, B.; van der Spoel, D. J. Mol. Model. 2001, 7,306.
28. [28]
  (28) Mackerell, A. D. J. Comput. Chem. 2004, 25, 1584.(28) Mackerell, A. D. J. Comput. Chem. 2004, 25, 1584.
29. [29]
  (29) Bussi, G.; Donadio, D.; Parrinello, M. J. Chem. Phys. 2007,126, 014101. doi: 10.1063/1.2408420(29) Bussi, G.; Donadio, D.; Parrinello, M. J. Chem. Phys. 2007,126, 014101. doi: 10.1063/1.2408420
30. [30]
  (30) Sayle, R.; Milnerwhite, E. Trends Biochem. Sci. 1995, 20, 374.doi: 10.1016/S0968-0004(00)89080-5(30) Sayle, R.; Milnerwhite, E. Trends Biochem. Sci. 1995, 20, 374.doi: 10.1016/S0968-0004(00)89080-5
31. [31]
  (31) Kabsch,W.; Sander, C. Biopolymers 1983, 22, 2577.(31) Kabsch,W.; Sander, C. Biopolymers 1983, 22, 2577.
32. [32]
  (32) Li,W. F.; Zhang, J.;Wang, J.;Wang,W. J. Am. Chem. Soc.2008, 130, 892. doi: 10.1021/ja075302g
  (32) Li,W. F.; Zhang, J.;Wang, J.;Wang,W. J. Am. Chem. Soc.2008, 130, 892. doi: 10.1021/ja075302g

扫一扫看文章

计量

PDF下载量: 504
文章访问数: 934
HTML全文浏览量: 6

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

静电排斥表面诱导溶菌酶分子站立

English

Standing Orientation of Lysozymes Induced by Electrostatically Repulsive Surface

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

静电排斥表面诱导溶菌酶分子站立

English

Standing Orientation of Lysozymes Induced by Electrostatically Repulsive Surface

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs