کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1926208 1536441 2010 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Evaluation of the conformational equilibrium of reduced hen egg lysozyme by antibodies to the native form
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Evaluation of the conformational equilibrium of reduced hen egg lysozyme by antibodies to the native form
چکیده انگلیسی

To evaluate the conformation of reduced HEL, the monoclonal antibodies HyC1 and HyC2, which recognize different conformational epitopes on native hen egg lysozyme (HEL), were used, and the kinetics of their interactions with native HEL, S-1,2-dicarboxyethylated HEL (DCE-HEL), and carboxymethylated Cys6 and Cys127 HEL (CM6,127-HEL) were assessed using surface plasmon resonance. Although their association rate constants differed 105-fold, their dissociation rate constants were essentially the same, suggesting that DCE-HEL and CM6,127-HEL possess conformations similar to that of native HEL when they bind antibodies. We considered that the ratio of the association rate constant of reduced HEL to native HEL represents the proportion of the native format determinant in equilibrium. Reduction of the Cys6–Cys127 disulfide bond would transform the epitope recognized by HyC1 into a non-native conformation similar to that of DCE-HEL. We show that monoclonal antibodies provide a sensitive tool for evaluation of the structural and hydrodynamic changes of proteins.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Archives of Biochemistry and Biophysics - Volume 494, Issue 2, 15 February 2010, Pages 145–150
نویسندگان
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