Conformational change and inactivation of arginine kinase in hexafluoroisopropanol: Evidence for existence of an unfolding equilibrium intermediate(精氨酸激酶在六氟异丙醇溶液中结构与活性的变化：去折叠平衡态中间体存在的证据)
利用内源荧光、1-苯氨基萘-8-磺酸(ANS)荧光、远紫外-圆二色光谱以及酶活力测定等手段研究了精氨酸激酶在1，1，1，3，3，3-六氟异丙醇(HFIP)溶液中变性后结构与活性的变化.为避免蛋白质聚沉的出现，实验中采用的变性剂浓度低于8%.当六氟异丙醇浓度为0.5%~1.5%时，蛋白质的二级结构和内源荧光发射峰位都没有发生明显的变化，而外源ANS荧光强度则显著增强.继续提高变性剂浓度至6%后，内源荧光发射峰位明显红移、二级结构含量增大、外源荧光减弱、酶活力完全丧失.内源荧光和外源ANS荧光的变化证明：在0.5%~1.5%六氟异丙醇浓度范围内，蛋白质分子以一种结构相对稳定并类似于天然蛋白结构的熔...
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| Format: | Article |
| Language: | zho |
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Zhejiang University Press
2007-09-01
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| Series: | Zhejiang Daxue xuebao. Lixue ban |
| Subjects: | |
| Online Access: | https://doi.org/zjup/1008-9497.2007.34.5.565-569 |
| _version_ | 1797236268357124096 |
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| author | LIHai-long(李海龙) MAYong(马勇) LIJie(李杰) SHIFeng(史锋) |
| author_facet | LIHai-long(李海龙) MAYong(马勇) LIJie(李杰) SHIFeng(史锋) |
| author_sort | LIHai-long(李海龙) |
| collection | DOAJ |
| description | 利用内源荧光、1-苯氨基萘-8-磺酸(ANS)荧光、远紫外-圆二色光谱以及酶活力测定等手段研究了精氨酸激酶在1，1，1，3，3，3-六氟异丙醇(HFIP)溶液中变性后结构与活性的变化.为避免蛋白质聚沉的出现，实验中采用的变性剂浓度低于8%.当六氟异丙醇浓度为0.5%~1.5%时，蛋白质的二级结构和内源荧光发射峰位都没有发生明显的变化，而外源ANS荧光强度则显著增强.继续提高变性剂浓度至6%后，内源荧光发射峰位明显红移、二级结构含量增大、外源荧光减弱、酶活力完全丧失.内源荧光和外源ANS荧光的变化证明：在0.5%~1.5%六氟异丙醇浓度范围内，蛋白质分子以一种结构相对稳定并类似于天然蛋白结构的熔球态中间体的状态存在.这种熔球态中间体也是精氨酸激酶在去折叠过程中出现的一个平衡态中间体. |
| first_indexed | 2024-04-24T17:01:09Z |
| format | Article |
| id | doaj.art-f16fc3f9dedf499b924b5cbbbb270182 |
| institution | Directory Open Access Journal |
| issn | 1008-9497 |
| language | zho |
| last_indexed | 2024-04-24T17:01:09Z |
| publishDate | 2007-09-01 |
| publisher | Zhejiang University Press |
| record_format | Article |
| series | Zhejiang Daxue xuebao. Lixue ban |
| spelling | doaj.art-f16fc3f9dedf499b924b5cbbbb2701822024-03-29T01:58:25ZzhoZhejiang University PressZhejiang Daxue xuebao. Lixue ban1008-94972007-09-01345565569zjup/1008-9497.2007.34.5.565-569Conformational change and inactivation of arginine kinase in hexafluoroisopropanol: Evidence for existence of an unfolding equilibrium intermediate(精氨酸激酶在六氟异丙醇溶液中结构与活性的变化：去折叠平衡态中间体存在的证据)LIHai-long(李海龙)0MAYong(马勇)1LIJie(李杰)2SHIFeng(史锋)3 1.College of Life Sciences, Zhejiang University, Hangzhou 310058, China( 1.浙江大学生命科学学院，浙江 杭州 310058) 1.College of Life Sciences, Zhejiang University, Hangzhou 310058, China( 1.浙江大学生命科学学院，浙江 杭州 310058) 2. Zhejiang Yangtze Delta Region Institute of Tsinghua University, Jiaxing 314000, China( 2.浙江清华长三角研究院，浙江 嘉兴 314000) 1.College of Life Sciences, Zhejiang University, Hangzhou 310058, China( 1.浙江大学生命科学学院，浙江 杭州 310058)利用内源荧光、1-苯氨基萘-8-磺酸(ANS)荧光、远紫外-圆二色光谱以及酶活力测定等手段研究了精氨酸激酶在1，1，1，3，3，3-六氟异丙醇(HFIP)溶液中变性后结构与活性的变化.为避免蛋白质聚沉的出现，实验中采用的变性剂浓度低于8%.当六氟异丙醇浓度为0.5%~1.5%时，蛋白质的二级结构和内源荧光发射峰位都没有发生明显的变化，而外源ANS荧光强度则显著增强.继续提高变性剂浓度至6%后，内源荧光发射峰位明显红移、二级结构含量增大、外源荧光减弱、酶活力完全丧失.内源荧光和外源ANS荧光的变化证明：在0.5%~1.5%六氟异丙醇浓度范围内，蛋白质分子以一种结构相对稳定并类似于天然蛋白结构的熔球态中间体的状态存在.这种熔球态中间体也是精氨酸激酶在去折叠过程中出现的一个平衡态中间体.https://doi.org/zjup/1008-9497.2007.34.5.565-569精氨酸激酶六氟异丙醇熔球态平衡态中间体 |
| spellingShingle | LIHai-long(李海龙) MAYong(马勇) LIJie(李杰) SHIFeng(史锋) Conformational change and inactivation of arginine kinase in hexafluoroisopropanol: Evidence for existence of an unfolding equilibrium intermediate(精氨酸激酶在六氟异丙醇溶液中结构与活性的变化：去折叠平衡态中间体存在的证据) Zhejiang Daxue xuebao. Lixue ban 精氨酸激酶 六氟异丙醇 熔球态 平衡态中间体 |
| title | Conformational change and inactivation of arginine kinase in hexafluoroisopropanol: Evidence for existence of an unfolding equilibrium intermediate(精氨酸激酶在六氟异丙醇溶液中结构与活性的变化：去折叠平衡态中间体存在的证据) |
| title_full | Conformational change and inactivation of arginine kinase in hexafluoroisopropanol: Evidence for existence of an unfolding equilibrium intermediate(精氨酸激酶在六氟异丙醇溶液中结构与活性的变化：去折叠平衡态中间体存在的证据) |
| title_fullStr | Conformational change and inactivation of arginine kinase in hexafluoroisopropanol: Evidence for existence of an unfolding equilibrium intermediate(精氨酸激酶在六氟异丙醇溶液中结构与活性的变化：去折叠平衡态中间体存在的证据) |
| title_full_unstemmed | Conformational change and inactivation of arginine kinase in hexafluoroisopropanol: Evidence for existence of an unfolding equilibrium intermediate(精氨酸激酶在六氟异丙醇溶液中结构与活性的变化：去折叠平衡态中间体存在的证据) |
| title_short | Conformational change and inactivation of arginine kinase in hexafluoroisopropanol: Evidence for existence of an unfolding equilibrium intermediate(精氨酸激酶在六氟异丙醇溶液中结构与活性的变化：去折叠平衡态中间体存在的证据) |
| title_sort | conformational change and inactivation of arginine kinase in hexafluoroisopropanol evidence for existence of an unfolding equilibrium intermediate 精氨酸激酶在六氟异丙醇溶液中结构与活性的变化 去折叠平衡态中间体存在的证据 |
| topic | 精氨酸激酶 六氟异丙醇 熔球态 平衡态中间体 |
| url | https://doi.org/zjup/1008-9497.2007.34.5.565-569 |
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