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Use of ion exchange chromatography for the study of RecA‐DNA interaction
Author(s) -
Takahashi Masayuki,
Hagmar Per
Publication year - 1991
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(91)80165-y
Subject(s) - cooperativity , dna , cooperative binding , chemistry , stoichiometry , monomer , molecule , ion chromatography , ion exchange , chromatography , dna binding protein , binding site , biophysics , crystallography , ion , biochemistry , biology , transcription factor , polymer , organic chemistry , gene
In vitro binding or 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 or 3 base pairs per RecA monomer. The binding exhibited cooperativity, and the stoichiometry suggested that RecA does not form complexes with two molecules or dsDNA. More than 90% of RecA molecules in the sample were active for DNA binding.