| 用多维NMR谱和圆二色CD光谱研究蛋白质亲环素A的结构稳定性 |
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| 引用本文： | 施燕红,林东海,黄剑英,沈旭. 用多维NMR谱和圆二色CD光谱研究蛋白质亲环素A的结构稳定性[J]. 中国化学, 2006, 24(7): 973-979. DOI: 10.1002/cjoc.200690184 |
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| 作者姓名： | 施燕红 林东海 黄剑英 沈旭 |
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| 作者单位： | Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 201203, China |
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| 基金项目： | Project supported by the Hundred Talent Project of Chinese Academy of Sciences and the National Natural Science Foundation of China （Nos 30470351, 30570352）.Acknowledgement The authors would like to thank Guo Hong-Xia and Cai Jian-Hua for their help in protein expression and purification. |
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| 摘 要： | The structural stability of cyclophilin A （CypA） was investigated using H/D exchange and temperature coefficients of chemical shifts of amide protons, monitored by 213 heteronuclear NMR spectroscopy. Amide proton exchange rates were measured by H/D exchange experiments for slow-exchange protons and measured by SEA （Solvent Exposed Amides）-HSQC experiments for fast-exchange protons. Temperature coefficients of chemical shifts and hydrogen exchange rates of amide protons show reasonably good correlation with the protein structure. Totally, 44 out of 153 non-proline assigned residues still exist in 86 d of hydrogen-deuterium exchange at 4 ℃, suggesting that CypA structure should be highly stable. Residues in secondary structures of α2, β1, β2, β5, β6 and β7 might constitute the hydrophobic core of the protein. The change in free energy of unfolding （ △Gu^H2O ） of CypA was estimated to be （21.99± 1.53） kJ·mol^-1 by circular dichroism （CD）. The large free energy change is also an indicator of the high structural stability.
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| 关 键 词： | 细胞亲环素A 结构稳定性 氢交换 NM 循环二色性 药物化学 |
| 收稿时间： | 2005-01-25 |
| 修稿时间： | 2005-01-252006-03-22 |
Study of Structural Stability of Cyclophilin A by NMR and Circular Dichroism Spectra |
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| SHI, Yan-Hong LIN, Dong-Hai HUANG, Jian-Ying SHEN, Xu. Study of Structural Stability of Cyclophilin A by NMR and Circular Dichroism Spectra[J]. Chinese Journal of Chemistry, 2006, 24(7): 973-979. DOI: 10.1002/cjoc.200690184 |
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| Authors: | SHI Yan-Hong LIN Dong-Hai HUANG Jian-Ying SHEN Xu |
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| Abstract: | The structural stability of cyclophilin A (CypA) was investigated using H/D exchange and temperature coefficients of chemical shifts of amide protons, monitored by 2D heteronuclear NMR spectroscopy. Amide proton exchange rates were measured by H/D exchange experiments for slow‐exchange protons and measured by SEA (Solvent Exposed Amides)‐HSQC experiments for fast‐exchange protons. Temperature coefficients of chemical shifts and hydrogen exchange rates of amide protons show reasonably good correlation with the protein structure. Totally, 44 out of 153 non‐proline assigned residues still exist in 86 d of hydrogen‐deuterium exchange at 4 °C, suggesting that CypA structure should be highly stable. Residues in secondary structures of α2, β1, β2, β5, β6 and β7 might constitute the hydrophobic core of the protein. The change in free energy of unfolding (ΔG u) of CypA was estimated to be (21.99±1.53) kJ·mol?1 by circular dichroism (CD). The large free energy change is also an indicator of the high structural stability. |
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| Keywords: | cyclophilin A structural stability hydrogen exchange NMR circular dichroism |
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