UV-induced ligand exchange in MHC class I protein crystals

J Am Chem Soc. 2009 Sep 2;131(34):12298-304. doi: 10.1021/ja9037559.

Abstract

High-throughput structure determination of protein-ligand complexes is central in drug development and structural proteomics. To facilitate such high-throughput structure determination we designed an induced replacement strategy. Crystals of a protein complex bound to a photosensitive ligand are exposed to UV light, inducing the departure of the bound ligand, allowing a new ligand to soak in. We exemplify the approach for a class of protein complexes that is especially recalcitrant to high-throughput strategies: the MHC class I proteins. We developed a UV-sensitive, "conditional", peptide ligand whose UV-induced cleavage in the crystals leads to the exchange of the low-affinity lytic fragments for full-length peptides introduced in the crystallant solution. This "in crystallo" exchange is monitored by the loss of seleno-methionine anomalous diffraction signal of the conditional peptide compared to the signal of labeled MHC beta2m subunit. This method has the potential to facilitate high-throughput crystallography in various protein families.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Crystallization
  • Crystallography, X-Ray
  • HLA-A2 Antigen / chemistry*
  • HLA-A2 Antigen / metabolism*
  • Humans
  • Ligands
  • Models, Molecular
  • Oligopeptides / chemistry*
  • Oligopeptides / metabolism*
  • Protein Binding / radiation effects
  • Protein Conformation / radiation effects
  • Selenium / chemistry
  • Ultraviolet Rays*

Substances

  • HLA-A2 Antigen
  • Ligands
  • Oligopeptides
  • Selenium