JAK2 JH2 Fluorescence Polarization Assay and Crystal Structures for Complexes with Three Small Molecules

  • ACS Med Chem Lett. 2017 May 17;8(6):614-617. doi: 10.1021/acsmedchemlett.7b00154.
Ana S Newton  1 ,  Luca Deiana  1 ,  David E Puleo  2 ,  José A Cisneros  1 ,  Kara J Cutrona  1 ,  Joseph Schlessinger  2 ,  William L Jorgensen  1
Affiliations
  • 1. Department of Chemistry, Yale University, New Haven, Connecticut 06520-8107, United States.
  • 2. Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut 06520-8066, United States.
PMID: 28626520 DOI: 10.1021/acsmedchemlett.7b00154
Abstract

A competitive fluorescence polarization (FP) assay is reported for determining binding affinities of probe molecules with the pseudokinase JAK2 JH2 allosteric site. The syntheses of the fluorescent 5 and 6 used in the assay are reported as well as Kd results for 10 compounds, including JNJ7706621, NVP-BSK805, and filgotinib (GLPG0634). X-ray crystal structures of JAK2 JH2 in complex with NVP-BSK805, filgotinib, and diaminopyrimidine 8 elucidate the binding poses.

Keywords
FP; Filgotinib (GLPG0634); Fluorescence polarization; JAK2; JAK2 JH2; JNJ7706621; Janus Kinase 2; NVP-BSK805; Protein crystallography; Pseudokinase domain.
Products