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

Ana S Newton; Luca Deiana; David E Puleo; José A Cisneros; Kara J Cutrona; Joseph Schlessinger; William L Jorgensen

doi:10.1021/acsmedchemlett.7b00154

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. eCollection 2017 Jun 8.

Authors

Ana S Newton¹, Luca Deiana¹, David E Puleo², José A Cisneros¹, Kara J Cutrona¹, Joseph Schlessinger², William L Jorgensen¹

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.

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 K_d 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.

Abstract

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