The high-resolution NMR structure of the early folding intermediate of the Thermus thermophilus ribonuclease H

J Mol Biol. 2008 Dec 12;384(2):531-9. doi: 10.1016/j.jmb.2008.09.044. Epub 2008 Sep 26.

Abstract

Elucidation of the high-resolution structures of folding intermediates is a necessary but difficult step toward the ultimate understanding of the mechanism of protein folding. Here, using hydrogen-exchange-directed protein engineering, we populated the folding intermediate of the Thermus thermophilus ribonuclease H, which forms before the rate-limiting transition state, by removing the unfolded regions of the intermediate, including an alpha-helix and two beta-strands (51 folded residues). Using multidimensional NMR, we solved the structure of this intermediate mimic to an atomic resolution (backbone rmsd, 0.51 A). It has a native-like backbone topology and shows some local deviations from the native structure, revealing that the structure of the folded region of an early folding intermediate can be as well defined as the native structure. The topological parameters calculated from the structures of the intermediate mimic and the native state predict that the intermediate should fold on a millisecond time scale or less and form much faster than the native state. Other factors that may lead to the slow folding of the native state and the accumulation of the intermediate before the rate-limiting transition state are also discussed.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Enzyme Stability
  • Hydrogen
  • Kinetics
  • Magnetic Resonance Spectroscopy
  • Molecular Mimicry
  • Protein Engineering
  • Protein Folding*
  • Protein Structure, Secondary
  • Ribonuclease H / chemistry*
  • Ribonuclease H / metabolism*
  • Thermus thermophilus / enzymology*
  • Time Factors

Substances

  • Hydrogen
  • Ribonuclease H

Associated data

  • PDB/2RPI