Kinetic and Thermodynamic Characterization of the R17 Coat Protein-Ribonucleic Acid Interaction

Jannette Carey, Olke C. Uhlenbeck*

*Corresponding author for this work

Research output: Contribution to journalArticle

108 Scopus citations

Abstract

A filter retention assay is used to examine the kinetic and equilibrium properties of the interaction between phage R17 coat protein and its 21-nucleotide RNA binding site. The kinetics of the reaction are consistent with the equilibrium association constant and indicate a diffusion-controlled reaction. The temperature dependence of Ka gives ΔH = −19 kcal/mol. This large favorable ΔH is partially offset by a ΔS = −30 cal mol−1 deg−1 to give a ΔG = −11 kcal/mol at 2 °C in 0.19 M salt. The binding reaction has a pH optimum centered around pH 8.5, but pH has no effect on ΔH. While the interaction is insensitive to the type of monovalent cation, the affinity decreases with the lyotropic series among monovalent anions. The ionic strength dependence of Ka reveals that ionic contacts contribute to the interaction. Most of the binding free energy, however, is a result of nonelectrostatic interactions.

Original languageEnglish (US)
Pages (from-to)2610-2615
Number of pages6
JournalBiochemistry
Volume22
Issue number11
DOIs
StatePublished - 1983

ASJC Scopus subject areas

  • Biochemistry

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