Thermodynamic analysis of a multifunctional RNA-binding protein, PhoRpp38, in the hyperthermophilic archaeon Pyrococcus horikoshii OT3

Biosci Biotechnol Biochem. 2012;76(6):1252-5. doi: 10.1271/bbb.120272. Epub 2012 Jun 7.

Abstract

The protein component PhoRpp38 of Pyrococcus horikoshii ribonuclease P (RNase P) is known to be a multifunctional RNA-binding protein. Previous biochemical data indicate that it binds to two stem-loops in RNase P RNA (PhopRNA). Thermodynamic analysis revealed that PhoRpp38 and PhopRNA interact with each other with an association constant (Ka) of 1.56×10(7) M(-1). It was further found that PhoRpp38 simultaneously binds two stem-loop structures in PhopRNA with approximately equal affinity. Crystals of PhoRpp38 in complex with the stem-loop were grown and diffracted to a resolution of 7.0 Å on a synchrotron X-ray source.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Archaeal Proteins / chemistry*
  • Archaeal Proteins / genetics
  • Archaeal Proteins / metabolism
  • Binding Sites
  • Calorimetry
  • Crystallography, X-Ray
  • Electrophoretic Mobility Shift Assay
  • Hot Temperature
  • Inverted Repeat Sequences
  • Kinetics
  • Molecular Sequence Data
  • Mutation
  • Protein Binding
  • Pyrococcus horikoshii / genetics
  • Pyrococcus horikoshii / metabolism*
  • RNA, Archaeal / chemistry*
  • RNA, Archaeal / genetics
  • RNA, Archaeal / metabolism
  • Ribonuclease P / chemistry*
  • Ribonuclease P / genetics
  • Ribonuclease P / metabolism
  • Thermodynamics

Substances

  • Archaeal Proteins
  • RNA, Archaeal
  • Ribonuclease P