Poly(ethylene oxide) facilitates the characterization of an affinity between strongly basic proteins with DNA by affinity capillary electrophoresis |
| |
Authors: | Tran Nguyet Thuy Taverna Myriam Miccoli Laurent Angulo Jaime F |
| |
Affiliation: | Groupe de Chimie Analytique Paris-Sud, EA 3343, Faculté de Pharmacie, Université Paris XI, F-92290 Chatenay-Malabry, France. thuy.tran@cep.u-psud.fr |
| |
Abstract: | In order to study kin17 protein-DNA affinity, we have developed a fast and reproducible capillary electrophoresis (CE) analysis of a strongly basic protein: kin17 protein, using a nonpermanent coating based on poly(ethylene oxide) (PEO) to avoid adsorption of kin17. The coating procedure was optimized to provide a residual and stable electroosmotic flow (EOF = 5 x 10(-5) cm(2)/V x s), exhibiting RSD of 0.3% and excellent long-term stability. Good intraday and interday reproducibility of kin17 migration times (0.8 and 0.3% relative standard deviation (RSD), respectively) enabled us to consider that the recovery percentage obtained for kin17 protein was satisfactory (79%). The potential of this PEO-based coating procedure was evaluated for affinity CE method in order to study the affinity of kin17 protein for two single-stranded DNA (ssDNA) models: polydeoxyadenylic acid and polydeoxycytidilic acid (pdA and pdC). Binding constants (1.5 x 10(7) +/- 17% and 1.7 x 10(7) + 25%M(-1)) were evaluated assuming a 1:1 affinity between kin17 and pdA or pdC, respectively. |
| |
Keywords: | Affinity capillary electrophoresis Higher order equilibria kin17 protein Protein adsorption |
本文献已被 PubMed 等数据库收录! |
|