Thermodynamic and activation parameters for binding of a pyrene-labeled substrate by the Tetrahymena ribozyme: docking is not diffusion-controlled and is driven by a favorable entropy change.
Association and dissociation rates for the pyrene-(pyr)-labeled oligoribonucleotide substrate pyrCUCU binding to the L-21 ScaI group I ribozyme are reported as a function of temperature. Combined with thermodynamic parameters for binding of pyrCUCU to rGGAGAA, the results allow calculation of the activation and thermodynamic parameters for docking of pyrCUCU into the catalytic core of the ribozyme. The activation enthalpy for docking is 22 kcal/mol, much larger than the approximately 4 kcal/mol expected for a diffusion-controlled process. Thus, docking is not diffusion-controlled. The activation and equilibrium entropies for docking are favorable at 21 and 37 eu, respectively. The results suggest the rate-limiting step and the driving force for docking may involve desolvation of RNA functional groups or of Mg2+ ions.
Li
Y
Y
Department of Chemistry, University of Rochester, New York 14627-0216, USA.
Bevilacqua
P C
PC
Mathews
D
D
Turner
D H
DH
D000818:Animals; D002384:Catalysis; D007700:Kinetics; D011721:Pyrenes; D016337:RNA, Catalytic / Q000378:metabolism*; D013768:Tetrahymena / Q000378:metabolism*; D013816:Thermodynamics
D011721:Pyrenes; D016337:RNA, Catalytic; C030984:pyrene