Use of ion exchange chromatography for the study of RecA-DNA interaction.
In vitro binding of RecA protein to double-stranded DNA (dsDNA) was studied using ion-exchange liquid chromatography. The method allowed quantification of both free DNA and free protein. The results unambiguously showed a binding stoichiometry of 3 base pairs per RecA monomer. The binding exhibited cooperativity, and the stoichiometry suggested that RecA does not form complexes with two molecules of dsDNA. More than 90% of RecA molecules in the sample were active for DNA binding.
Takahashi
M
M
Institut de Biologie Molécule et Cellulaire du CNRS, Strasbourg, France.
Hagmar
P
P
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D011485:Protein Binding; D011938:Rec A Recombinases / Q000378:metabolism*
D004268:DNA-Binding Proteins; C022571:adenosine 5'-O-(3-thiotriphosphate); D000255:Adenosine Triphosphate; D004247:DNA; D011938:Rec A Recombinases