title: Interaction of Escherichia coli RecA protein with ATP and its analogues.
abstract: Interactions of Escherichia coli RecA protein with ATP and its analogues in the absence of DNA were studied by circular dichroic (CD) spectroscopy. The binding of RecA protein to ATP increased the CD band of ATP at around 260 nm. The positive CD band of the RecA protein-ATP complex suggested that the bound ATP was in the anti conformation, in accord with X-ray crystallographic data [Story, R.M. and Steitz, T.A. (1992) Nature 355, 374-376]. At pH 7.5 and at 25 degrees C the dissociation constant (Kd) and thermodynamic parameters for the binding of ATP to RecA protein were 18 microM (delta G = -6.5 kcal.mol-1), delta H = 0 kcal.mol-1, and delta S = 22 cal.mol-1.K-1. A non-hydrolyzable ATP analogue, adenosine 5'-O-(3-thiotriphosphate) (ATP gamma S), gave a spectral change similar to that of ATP. The Kd for this analogue, 22 microM, was very close to the Km of ATP. These results in the absence of single-stranded DNA were different from those obtained by kinetic analysis [Weinstock, G.M. et al. (1981) J. Biol. Chem. 256, 8850-8855], which indicated that the inhibition constant of ATP gamma S was much smaller than the Km of ATP in the presence of DNA. For other ATP analogues (dATP, ADP, and dADP), similar spectral changes were observed, and their Kd values ranged from 19 to 54 microM. UTP, dUTP, and TTP also gave CD spectral changes, but not AMP, GTP, dGTP, CTP, and dCTP.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Biology, Faculty of Science, Osaka University.
authors.forename: R
authors.initials: R
authors.lastname: Watanabe
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: Takamatsu
authors.forename: R
authors.initials: R
authors.lastname: Kato
authors.forename: S
authors.initials: S
authors.lastname: Kuramitsu
mesh_terms: D000255:Adenosine Triphosphate / Q000037:antagonists & inhibitors / Q000378:metabolism*; D001665:Binding Sites; D002942:Circular Dichroism; D004926:Escherichia coli / Q000378:metabolism; D007700:Kinetics; D011938:Rec A Recombinases / Q000378:metabolism*; D012265:Ribonucleotides / Q000378:metabolism*
chemical_list: D012265:Ribonucleotides; D000255:Adenosine Triphosphate; D011938:Rec A Recombinases