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Spectral studies on the conformational transitions of bovine insulin during denaturant-induced unfolding

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Abstract

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.

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Authors and Affiliations

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

    Xu Ji, Xiaojuan Ma & Liujiao Bian

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  1. Xu Ji
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  2. Xiaojuan Ma
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  3. Liujiao Bian
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Correspondence to Liujiao Bian.

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Supported by the National Natural Science Foundation of China(No.21075097).

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Ji, X., Ma, X. & Bian, L. Spectral studies on the conformational transitions of bovine insulin during denaturant-induced unfolding. Chem. Res. Chin. Univ. 30, 222–227 (2014). https://doi.org/10.1007/s40242-014-3372-z

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  • DOI: https://doi.org/10.1007/s40242-014-3372-z

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