title: Local folding of the N-terminal domain of Escherichia coli RecA controls protein-protein interaction.
abstract: To obtain structural information about the self-association of the protein RecA, we studied urea denaturation of RecA by circular dichroism spectroscopy and gel filtration. Gel filtration analysis showed that urea at low concentrations, 1.0-1.2 M, dissociated the RecA oligomer to almost a monomeric state prior to the unfolding of each molecule. Upon treatment with 1.0 M urea, the circular dichroism spectrum showed a decrease in the alpha-helical content of RecA. A similar decrease was observed in the absence of urea for RecA at an extremely low protein concentration; the RecA oligomer dissociated to an almost completely monomeric state. The properties of RecA at low urea concentrations were similar to those of a truncated RecA lacking the first 33 N-terminal residues (Delta33RecA). Addition of a synthetic peptide corresponding to the 33 N-terminal residues to Delta33RecA increased the alpha-helical content. These results suggest that local folding of the N-terminal domain is coupled to protein-protein interactions of monomeric RecA, which are involved in the regulation of filament formation. The dissociation constant for interaction between RecA monomers was determined from the ellipticity data to be 0.1 microM.
authors.affiliation: Department of Biology, Graduate School of Science, Osaka University, Toyonaka, Osaka 560, Japan.
authors.forename: R
authors.initials: R
authors.lastname: Masui
authors.forename: T
authors.initials: T
authors.lastname: Mikawa
authors.forename: S
authors.initials: S
authors.lastname: Kuramitsu
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D002850:Chromatography, Gel; D002942:Circular Dichroism; D004926:Escherichia coli / Q000378:metabolism*; D011485:Protein Binding; D017510:Protein Folding; D011938:Rec A Recombinases / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet; D014508:Urea / Q000737:chemistry
chemical_list: D000255:Adenosine Triphosphate; D014508:Urea; D011938:Rec A Recombinases