کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1174804 961769 2011 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Evaluation of dissociation constants from competition binding experiments based on the relative binding ratio
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Evaluation of dissociation constants from competition binding experiments based on the relative binding ratio
چکیده انگلیسی

Methods probing protein–DNA associations include direct binding titrations and competition binding experiments. For the latter, we present here a simple procedure allowing the quantitative evaluation of dissociation constants. We show that the ratio between the fraction of a DNA probe bound to protein in the absence of competitor and that in the presence of competitor is, at large competitor concentrations, a linear function of the competitor concentration, and we derive equations allowing the dissociation constant for the protein–competitor complex to be evaluated from the slope. We show further that a self-competition experiment, where the DNA probe and competitor are chemically the same species, can be used as a complement to a direct titration to determine the fraction of protein that is correctly folded for specific DNA binding. Thus, such a combination of direct and self-competition titration can be used as a check of the conformational purity of DNA binding proteins.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 409, Issue 1, 1 February 2011, Pages 66–73
نویسندگان
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