Kinetic and thermodynamic analysis of RNA-protein interactions in the RNase P holoenzyme from escherichia colf

Simon J. Talbot, Sidney Altman

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

A gel retardation assay has been used to examine the kinetic and equilibrium properties of the interaction between C5 protein and M1 RNA in the formation of the ribonuclease P holoenzyme from Escherichia coli. The interaction is relatively insensitive to the identity of the monovalent anions present and to pH in the range 7.0-9.0, but it has a more critical requirement for specific monovalent and divalent cations: NH4+, K+, Mg2+, Ca2+, and Mn2+ all promote efficient formation of the complex. A positive ΔS (+6.4 cal mol-1 deg-1) and a negative ΔH (-11.3 kcal mol-1) combine to give a ΔG equal to -13.3 kcal mol-1 at 37 °C in 0.42 M salt. The binding reaction is sensitive to the concentration of monovalent and divalent cations, with the affinity increasing with increasing ionic strength (δ log Ka/δ log [NH4+] = +2.7 ± 0.1). The dependence of Kd on the ionic strength and the positive ΔS suggests that hydrophobic and stacking interactions contribute significantly to the formation of the RNase P holoenzyme.

Original languageEnglish (US)
Pages (from-to)1406-1411
Number of pages6
JournalBiochemistry
Volume33
Issue number6
StatePublished - 1994
Externally publishedYes

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Ribonuclease P
Escherichia
Monovalent Cations
Holoenzymes
Divalent Cations
Ionic strength
Thermodynamics
Osmolar Concentration
RNA
Kinetics
Electrophoretic Mobility Shift Assay
Hydrophobic and Hydrophilic Interactions
Escherichia coli
Anions
Assays
Proteins
Salts
Gels

ASJC Scopus subject areas

  • Biochemistry

Cite this

Kinetic and thermodynamic analysis of RNA-protein interactions in the RNase P holoenzyme from escherichia colf. / Talbot, Simon J.; Altman, Sidney.

In: Biochemistry, Vol. 33, No. 6, 1994, p. 1406-1411.

Research output: Contribution to journalArticle

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