Predictions of CCR1 chemokine receptor structure and BX 471 antagonist binding followed by experimental validation

J Biol Chem. 2006 Sep 15;281(37):27613-20. doi: 10.1074/jbc.M601389200. Epub 2006 Jul 12.

Abstract

A major challenge in the application of structure-based drug design methods to proteins belonging to the superfamily of G protein-coupled receptors (GPCRs) is the paucity of structural information (1). The 19 chemokine receptors, belonging to the Class A family of GPCRs, are important drug targets not only for autoimmune diseases like multiple sclerosis but also for the blockade of human immunodeficiency virus type 1 entry (2). Using the MembStruk computational method (3), we predicted the three-dimensional structure of the human CCR1 receptor. In addition, we predicted the binding site of the small molecule CCR1 antagonist BX 471, which is currently in Phase II clinical trials (4). Based on the predicted antagonist binding site we designed 17 point mutants of CCR1 to validate the predictions. Subsequent competitive ligand binding and chemotaxis experiments with these mutants gave an excellent correlation to these predictions. In particular, we find that Tyr-113 and Tyr-114 on transmembrane domain 3 and Ile-259 on transmembrane 6 contribute significantly to the binding of BX 471. Finally, we used the predicted and validated structure of CCR1 in a virtual screening validation of the Maybridge data base, seeded with selective CCR1 antagonists. The screen identified 63% of CCR1 antagonists in the top 5% of the hits. Our results indicate that rational drug design for GPCR targets is a feasible approach.

MeSH terms

  • Animals
  • Binding Sites
  • Chemotaxis
  • Epitopes / chemistry
  • Humans
  • Mice
  • Models, Molecular
  • Mutation
  • Phenylurea Compounds / antagonists & inhibitors*
  • Phenylurea Compounds / pharmacology*
  • Piperidines / antagonists & inhibitors*
  • Piperidines / pharmacology*
  • Protein Binding
  • Protein Conformation
  • Protein Structure, Tertiary
  • Receptors, CCR1
  • Receptors, Chemokine / chemistry*
  • Software
  • Tyrosine / chemistry

Substances

  • CCR1 protein, human
  • Ccr1 protein, mouse
  • Epitopes
  • Phenylurea Compounds
  • Piperidines
  • Receptors, CCR1
  • Receptors, Chemokine
  • Tyrosine
  • BX 471