Investigation of ligand interactions with human RXRalpha by hydrogen/deuterium exchange and mass spectrometry

J Am Soc Mass Spectrom. 2006 Nov;17(11):1510-7. doi: 10.1016/j.jasms.2006.05.016. Epub 2006 Jul 26.

Abstract

Several different agonists of the retinoic X receptor alpha (hRXRalpha) were examined for their effects on the amide H/D exchange kinetics of the homodimeric protein using mass spectrometry. Some agonists, LG 100268, SR11246, and DHA, bind such that slower deuterium exchange-in occurs compared with 9-cis-retinoic acid (9-cis-RA), whereas others, fenretinide and methoprenic acid, result in poorer protection during binding and hence faster exchange-in. Protection against H/D exchange by different agonists and the inhibition of H/D exchange kinetics relative to 9-cis-RA varies markedly in different regions of the protein. Agonists LG 100268, SR11246, and DHA generally inhibit faster exchange processes in the ligand binding regions of hRXRalpha than does the native ligand 9-cis-RA. In at least half of these regions, the level of protection by 9-cis-RA lags behind the agonists even after 60 min. Methoprenic acid did not significantly protect hRXRalpha against amide hydrogen exchange. An efficient method is described for comparing the effects of different agonists on the protein structure of the agonist-RXRalpha complex.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Deuterium Exchange Measurement / methods*
  • Humans
  • Hydrogen / chemistry*
  • Ligands
  • Molecular Sequence Data
  • Protein Conformation*
  • Retinoid X Receptor alpha / chemistry*
  • Spectrometry, Mass, Electrospray Ionization*

Substances

  • Ligands
  • Retinoid X Receptor alpha
  • Hydrogen