title: Coupling between substrate binding and allosteric regulation in ribozyme catalysis.
abstract: The contribution of substrate binding to allosteric regulation in the ribozyme catalysis has been investigated using allosteric ribozymes consisting of the hammerhead ribozyme and a flavin mononucleotide (FMN) aptamer. Kinetic parameters were measured for various lengths of the substrates with a wide range of binding energy. The maximum cleavage rate of each ribozyme was retained with the long substrates. However, the cleavage rates largely decreased by the truncation of the substrates according to loss in the free energy of substrate binding. The high sensitivity to the substrate lengths is attributed to the increase in the energetic requirement for the catalytic core folding, which is caused by the incorporation of the aptamer region. One role of FMN binding is assisting the promotion of the core folding through the stabilization of the aptamer domain. The allosteric effect is significantly expressed only when the substrate binding energy is insufficient for the core folding of the ribozyme-substrate complex. This type of allosteric interaction dominates the substrate dependency of another type of regulation. These results demonstrate that an adequate correlation between the type of regulation and the substrate binding is responsible for the effective allosteric interaction in the kinetic process.
authors.affiliation: Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Araki
authors.forename: M
authors.initials: M
authors.lastname: Hashima
authors.forename: Y
authors.initials: Y
authors.lastname: Okuno
authors.forename: Y
authors.initials: Y
authors.lastname: Sugiura
mesh_terms: D000494:Allosteric Regulation / Q000502:physiology; D001483:Base Sequence / Q000235:genetics / Q000502:physiology; D001665:Binding Sites / Q000502:physiology; D004734:Energy Metabolism / Q000502:physiology*; D005486:Flavin Mononucleotide / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D017510:Protein Folding; D016337:RNA, Catalytic / Q000378:metabolism*; D013379:Substrate Specificity / Q000502:physiology
chemical_list: D016337:RNA, Catalytic; C427204:hammerhead ribozyme; D005486:Flavin Mononucleotide