Evaluation of dissociation constants from competition binding experiments based on the relative binding ratio

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F11%3A00054323" target="_blank" >RIV/00216224:14310/11:00054323 - isvavai.cz</a>

Result on the web

<a href="http://dx.doi.org/10.1016/j.ab.2010.09.023" target="_blank" >http://dx.doi.org/10.1016/j.ab.2010.09.023</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.ab.2010.09.023" target="_blank" >10.1016/j.ab.2010.09.023</a>

Result language

angličtina

Original language name

Evaluation of dissociation constants from competition binding experiments based on the relative binding ratio

Original language description

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 selfcompetition 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.

Czech name

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Czech description

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Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

CB - Analytical chemistry, separation

OECD FORD branch

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Project

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Continuities

S - Specificky vyzkum na vysokych skolach

Publication year

2011

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Analytical Biochemistry

ISSN

0003-2697

e-ISSN

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Volume of the periodical

409

Issue of the periodical within the volume

1

Country of publishing house

CZ - CZECH REPUBLIC

Number of pages

8

Pages from-to

66-73

UT code for WoS article

000286126500010

EID of the result in the Scopus database

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