Guanidine hydrochloride and urea-induced unfolding of Brugia malayi hexokinase

被引：16

作者：

Singh, Alok Ranjan ^{[1
]}

Joshi, Shweta ^{[1
]}

Arya, Rahul ^{[2
]}

Kayastha, Arvind Mohan ^{[3
]}

Saxena, Jitendra Kumar ^{[1
]}

机构：

[1] Cent Drug Res Inst, Div Biochem, Lucknow 226001, Uttar Pradesh, India

[2] Int Ctr Genet Engn & Biotechnol, Virol Grp, New Delhi 110067, India

[3] Banaras Hindu Univ, Sch Biotechnol, Varanasi 221005, Uttar Pradesh, India

来源：

EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS | 2010年 / 39卷 / 02期

关键词：

Guanidine hydrochloride; Urea denaturation; Multimeric protein; Dimer; BmHk; YEAST HEXOKINASE; MOLECULAR-CLONING; CRYSTAL-STRUCTURE; PROTEIN; GLUCOSE; STABILITY; REFINEMENT; INSIGHTS; CHLORIDE; FORMS;

D O I：

10.1007/s00249-009-0539-5

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

Guanidine hydrochloride and urea-induced unfolding of B. malayi hexokinase (BmHk), a tetrameric protein, was examined in detail by using various optical spectroscopic techniques, enzymatic activity measurements, and size-exclusion chromatography. The equilibrium unfolding of BmHk by guanidine hydrochloride (GdmCl) and urea proceeded through stabilization of several unique oligomeric intermediates. In the presence of low concentrations of GdmCl, stabilization of an enzymatically active folded dimer of BmHk was observed. However an enzymatically inactive dimer of BmHk was observed for urea-treated BmHk. This is the first report of an enzymatically active dimer of hexokinase from any human filarial parasite. Furthermore, although complete recovery of the native enzyme was observed on refolding of BmHk samples denatured by use of low concentrations of GdmCl or urea, no recovery of the native enzyme was observed for BmHk samples denatured by use of high concentrations of GdmCl or urea.

引用

页码：289 / 297

页数：9

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