Exploring protein interiors: The role of a buried histidine in the KH module fold

Franca Fraternali, Pietro Amodeo, Giovanna Musco, Michael Nilges, Annalisa Pastore

Research output: Contribution to journalArticlepeer-review

Abstract

The K-homology (KH) module is a novel RNA-binding motif. The structures of a representative KH motif from vigilin (vig-KH6) and of the first KH domain of fmr1 have been recently solved by nuclear magnetic resonance (NMR) and automated assignment-refinement techniques (ARIA). While a hydrophobic residue is found at position 21 in most of the KH modules, a buried His is conserved in all the 15 KH repeats of vigilin. This position must therefore have a key structural role in stabilizing the hydrophobic core. In the present work, we have addressed the following questions in order to obtain a detailed description of the role of His 21:i) what is the exact role of the histidine in the hydrophobic core of vig-KH6? ii) can we define the interactions that allow a conserved buried position to be occupied by a histidine both in vig-KH6 and in the whole vigilin KH sub-family? iii) how is the structure and stability of vig-KH6 influenced by the state of protonation of this histidine? To answer these questions, we have carried out an extensive refinement of the vig-KH6 structure using both an improved ARIA protocol starting from different initial structures and successively running restrained and unrestrained trajectories in water. An analysis of the stability of secondary structural elements, solvent accessibility, and hydrogen bonding patterns allows hypothesis on the structural role of residue His 21 and on the interactions that this residue forms with the environment. The importance of the protonation state of His 21 on the stability of the KH fold was addressed and validated by experimental results.

Original languageEnglish
Pages (from-to)484-496
Number of pages13
JournalProteins: Structure, Function and Genetics
Volume34
Issue number4
DOIs
Publication statusPublished - Mar 1 1999

Keywords

  • Fmr1
  • Fragile X
  • Molecular dynamics
  • Stability
  • Vigilin

ASJC Scopus subject areas

  • Genetics
  • Structural Biology
  • Biochemistry

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