双糖保护剂对胰岛素活性结构稳定性的影响
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摘要
采用微量热分析法并结合分子动力学模拟,研究了不同双糖保护剂对胰岛素的保护效果.结果表明,通过区域选择性吸附,双糖保护剂可以在胰岛素表面产生保护作用,提高胰岛素的热稳定性.在4种双糖保护剂中,吸附作用越强的保护剂对胰岛素热稳定性的提升效果越好.随着保护剂质量分数的增加,胰岛素的热稳定性呈先增强后稳定的趋势.进一步分析了保护剂与水分子之间的相互作用,得出双糖保护剂与胰岛蛋白之间的直接相互作用可能是提高其热稳定性的主要原因.
The approaches of microcalorimetry and molecular dynamic simulation were adopted to investigate the protective effect caused by adding protectants.The results indicate that disaccharide protectants are able to maintain the molecular structure of insulin and contribute to the thermal stability of insulin due to the regional selective adsorption of protectants on the surface of insulin.Among four disaccharide protectants,the protectant with stronger adsorption ability conducts better performance in stabilizing the molecular structure of protein.With increasing the concentration of protectants,the thermal stability of insulin changes from increasing to gradually stable state.A further analysis upon the interaction between protectants and water reflects that increasing the thermal stability of insulin is attributed mainly to the interaction between insulin and disaccharides.
The approaches of microcalorimetry and molecular dynamic simulation were adopted to investigate the protective effect caused by adding protectants.The results indicate that disaccharide protectants are able to maintain the molecular structure of insulin and contribute to the thermal stability of insulin due to the regional selective adsorption of protectants on the surface of insulin.Among four disaccharide protectants,the protectant with stronger adsorption ability conducts better performance in stabilizing the molecular structure of protein.With increasing the concentration of protectants,the thermal stability of insulin changes from increasing to gradually stable state.A further analysis upon the interaction between protectants and water reflects that increasing the thermal stability of insulin is attributed mainly to the interaction between insulin and disaccharides.
引文
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[5]RICHTER B,NEISES G.Human insulin versus animal insulin in people with diabetes mellitus[J].Cochrane Database of Systematic Reviews,2003(3):1469-1493.
[6]GILL P,MOGHADAM T T,RANJBAR B.Differential scanning calorimetry techniques:applications in biology and nanoscience[J].Journal of Biomolecular Techniques,2010,21(4):167-193.
[7]KRGER P,STRASSBURGER W,WOLLMER A,et al.The simulated dynamics of the insulin monomer and their relationship to the molecule’s structure[J].European Biophysics Journal,1987,14(8):449-459.
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[9]CUETO M,DORTA M J,MUNGUA O,et al.New approach to stability assessment of protein solution formulations by differential scanning calorimetry[J].International Journal of Pharmaceutics,2003,252(1/2):159-166.
[10]KRISHNA N R S,PATTABHI V,RAJAN S S.Metal induced conformational changes in human insulin:crystal structures of Sr+2,Ni+2and Cu+2complexes of human insulin[J].Protein&Peptide Letters,2011,18(5):457-466.
[11]DEGTYARENKO K,DE MATOS P,ENNIS M,et al.ChEBI:a database and ontology for chemical entities of biological interest[J].Nucleic Acids Research,2008,36(1):344-350.
[12]李代禧,刘亿淑,张建民,等.扑热息痛与β-环糊精包合物的热分解动力学和包合结构的研究[J].上海理工大学学报,2007,29(4):373-377.
[13]ABRAHAM M J,MURTOLA T,SCHULZ R,et al.GROMACS:high performance molecular simulations through multi-level parallelism from laptops to supercomputers[J].Softwarex,2015,1:19-25.
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[15]HORNAK V,ABEL R,OKUR A,et al.Comparison of multiple Amber force fields and development of improved protein backbone parameters[J].Proteins:Structure Function,and Bioinformatics,2006,65(3):712-725.
[16]ABASCAL J L F,VEGA C.A general purpose model for the condensed phases of water:TIP4P/2005[J].The Journal of Chemical Physics,2005,123(23):234505.
[17]VAN DER SPOEL D,VAN MAAREN P J.The origin of layer structure artifacts in simulations of liquid water[J].Journal of Chemical Theory and Computation,2006,2(1):1-11.
[18]DARDEN T,YORK D,PEDERSEN L.Particle mesh ewald:an N-log(N)method for ewald sums in large systems[J].The Journal of Chemical Physics,1993,98(12):10089.
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[21]RENTENIER A,MORETTO-CAPELLE P,BORDENAVEMONTESQUIEU D,et al.Analysis of fragment size distributions in collisions of monocharged ions with the C60 molecule[J].Journal of Physics B:Atomic,Molecular and Optical Physics,2005,38(7):789-806.
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[24]TAREK M,TOBIAS D J.Single-particle and collective dynamics of protein hydration water:a molecular dynamics study[J].Physical Review Letters,2002,89(27):275501.
[25]DASTIDAR S G,MUKHOPADHYAY C.Structure,dynamics,and energetics of water at the surface of a small globular protein:a molecular dynamics simulation[J].Physical Review E,2003,68(2):021921.