NMR backbone assignments of the tyrosine kinase domain of human fibroblast growth factor receptor 3 in apo state and in complex with inhibitor PD173074

Abstract

Fibroblast growth factors receptors (FGFR) are transmembrane protein tyrosine kinases involved in many cellular process, including growth, differentiation and angiogenesis. Dysregulation of FGFR enzymatic activity is associated with developmental disorders and cancers; therefore FGFRs have become attractive targets for drug discovery, with a number of agents in late-stage clinical trials. Here, we present the backbone resonance assignments of FGFR3 tyrosine kinase domain in the ligand-free form and in complex with the canonical FGFR kinase inhibitor PD173074. Analysis of chemical shift changes upon inhibitor binding highlights a characteristic pattern of allosteric network perturbations that is of relevance for future drug discovery activities aimed at development of conformationally-selective FGFR inhibitors.

NMR Resonance Assignment of the LA Motif of Human LA-Related Protein 1

“…Uniformly [ 13 C, 15 N]-labeled His6-TEV-HsLARP1(392-486) was expressed using M9 minimal medium containing 1 g/L 15 NH4Cl, 2 g/L 13 C-D-glucose, and 40 µg/mL kanamycin. Uniformly were expressed using protocols published previously [34,35]. Uniformly [ 15 N, 2-13 C-glycerol]-labeled and [ 15 N, 1,3-13 C-glycerol]-labeled His6-TEV-HsLARP1(392-486) were expressed using protocols published previously [36][37][38].…”

Backbone resonance assignments of the catalytic and regulatory domains of Ca2+/calmodulin-dependent protein kinase 1D

“…However, protein kinases are challenging to study by NMR, often yielding poor spectra due to their intrinsic dynamics and large molecular weights. Thus few protein kinases have been assigned (Langer et al 2004 ; Vogtherr et al 2005 , 2006 ; Gelev et al 2006 ; Vajpai et al 2008 ; Masterson et al 2009 ; Xiao et al 2015 ; Serimbetov et al 2017 ; Sanfelice et al 2018 ). Following optimization of solution conditions, the autoinhibited human CaMK1D was found to be amenable to NMR analysis, yielding resolved spectra under physiological buffer conditions.…”