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...
Main Authors: | LIHai-long(李海龙), MAYong(马勇), LIJie(李杰), SHIFeng(史锋) |
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Format: | Article |
Language: | zho |
Published: |
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 |
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