Spectral Studies on the Conformational Transitions of Bovine Insulin During Denaturant-induced Unfolding

doi:10.1007/s40242-014-3372-z

高等学校化学研究 ›› 2014, Vol. 30 ›› Issue (2): 222-227.doi: 10.1007/s40242-014-3372-z

Spectral Studies on the Conformational Transitions of Bovine Insulin During Denaturant-induced Unfolding

JI Xu, MA Xiaojuan, BIAN Liujiao

College of Life Science, Northwest University, Xi'an 710069, P. R. China

收稿日期:2013-09-05 修回日期:2013-11-26 出版日期:2014-04-01 发布日期:2013-12-04
通讯作者: BIAN Liujiao E-mail:bianliujiao@sohu.com
基金资助:
Supported by the National Natural Science Foundation of China(No.21075097).

Spectral Studies on the Conformational Transitions of Bovine Insulin During Denaturant-induced Unfolding

JI Xu, MA Xiaojuan, BIAN Liujiao

College of Life Science, Northwest University, Xi'an 710069, P. R. China

Received:2013-09-05 Revised:2013-11-26 Online:2014-04-01 Published:2013-12-04
Contact: BIAN Liujiao E-mail:bianliujiao@sohu.com
Supported by:
Supported by the National Natural Science Foundation of China(No.21075097).

摘要/Abstract

摘要：

Transitions among various molecule states and conformational changes of bovine insulin were investigated under different denaturing conditions by means of fluorescence phase diagrams, fluorescence quenching, 1-anilinonaphthalene-8-sulfonate(ANS) binding assay and circular dichroism(CD) spectra. In both guanidine hydrochloride(GuHCl)-and urea-denatured procedures, the spatial structure of insulin molecules changed from ordered states to relative unordered ones with the increasing of denaturant concentration. The GuHCl-denatured process followed a four-state model, for there were two intermediates existed in 2.0 and 6.0 mol/L GuHCl, respectively. Intermediate I₁ is more compact than the normal protein. And intermediate I₂ has lost most of the secondary structures. When GuHCl concentration was above 6.0 mol/L, the fluorophores originally existed in the internal of insulin molecules would expose to the surface. However, the urea-denatured process followed a three-state model, only one intermediate existed in 2.5 mol/L urea. During the urea-denatured procedure, the fluorophores originally existed in the internal of insulin molecules didn't expose to the surface.

关键词: Bovine insulin, Unfolding, Intermediate, Guanidine hydrochloride, Urea

Abstract:

Key words: Bovine insulin, Unfolding, Intermediate, Guanidine hydrochloride, Urea

JI Xu, MA Xiaojuan, BIAN Liujiao. Spectral Studies on the Conformational Transitions of Bovine Insulin During Denaturant-induced Unfolding[J]. 高等学校化学研究, 2014, 30(2): 222-227.

JI Xu, MA Xiaojuan, BIAN Liujiao. Spectral Studies on the Conformational Transitions of Bovine Insulin During Denaturant-induced Unfolding[J]. Chemical Research in Chinese Universities, 2014, 30(2): 222-227.

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Spectral Studies on the Conformational Transitions of Bovine Insulin During Denaturant-induced Unfolding

Spectral Studies on the Conformational Transitions of Bovine Insulin During Denaturant-induced Unfolding

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