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

J 2011

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

KOZELKA, Jiří

Základní údaje

Originální název

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

Autoři

KOZELKA, Jiří

Vydání

Analytical Biochemistry, San Diego, Academic Press, 2011, 0003-2697

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 2.996

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/11:00054323

Organizační jednotka

Přírodovědecká fakulta

DOI

https://doi.org/10.1016/j.ab.2010.09.023

UT WoS

000286126500010

Klíčová slova anglicky

Protein–DNA interactions Electrophoretic mobility shift assay Competition binding experiment

Štítky

AKR, rivok

Změněno: 4. 1. 2012 16:55, prof. Dr. Jiří Kozelka, PhD.

Anotace

V originále

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.

Citovat

KOZELKA, Jiří. Evaluation of dissociation constants from competition binding experiments based on the relative binding ratio. Analytical Biochemistry. San Diego: Academic Press, 2011, roč. 409, č. 1, s. 66-73. ISSN 0003-2697. Dostupné z: https://doi.org/10.1016/j.ab.2010.09.023.

@article{961402,
   author = {Kozelka, Jiří},
   article_location = {San Diego},
   article_number = {1},
   doi = {https://doi.org/10.1016/j.ab.2010.09.023},
   keywords = {Protein–DNA interactions Electrophoretic mobility shift assay Competition binding experiment},
   language = {eng},
   issn = {0003-2697},
   journal = {Analytical Biochemistry},
   title = {Evaluation of dissociation constants from competition binding experiments based on the relative binding ratio},
   volume = {409},
   year = {2011}
}

TY  - JOUR
ID  - 961402
AU  - Kozelka, Jiří
PY  - 2011
TI  - Evaluation of dissociation constants from competition binding experiments based on the relative binding ratio
JF  - Analytical Biochemistry
VL  - 409
IS  - 1
SP  - 66-73
EP  - 66-73
PB  - Academic Press
SN  - 00032697
KW  - Protein–DNA interactions Electrophoretic mobility shift assay Competition binding experiment
N2  - 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.
ER  -

KOZELKA, Jiří. Evaluation of dissociation constants from competition binding experiments based on the relative binding ratio. \textit{Analytical Biochemistry}. San Diego: Academic Press, 2011, roč.~409, č.~1, s.~66-73. ISSN~0003-2697. Dostupné z: https://doi.org/10.1016/j.ab.2010.09.023.

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