Family-wide Structural Analysis of Human Numb-Associated Protein Kinases

Abstract

SummaryThe highly diverse Numb-associated kinase (NAK) family has been linked to broad cellular functions including receptor-mediated endocytosis, Notch pathway modulation, osteoblast differentiation, and dendrite morphogenesis. Consequently, NAK kinases play a key role in a diverse range of diseases from Parkinson's and prostate cancer to HIV. Due to the plasticity of this kinase family, NAK kinases are often inhibited by approved or investigational drugs and have been associated with side effects, but they are also potential drug targets. The presence of cysteine residues in some NAK family members provides the possibility for selective targeting via covalent inhibition. Here we report the first high-resolution structures of kinases AAK1 and BIKE in complex with two drug candidates. The presented data allow a comprehensive structural characterization of the NAK kinase family and provide the basis for rational design of selective NAK inhibitors.

Baricitinib as potential treatment for 2019-nCoV acute respiratory disease

“…By contrast, one of the six highaffinity AAK1-binding drugs was the janus kinase inhibitor baricitinib, which also binds the cyclin G-associated kinase, another regulator of endocytosis. 6 Because the plasma concentration of baricitinib on therapeutic dosing (either as 2 mg or 4 mg once daily) is sufficient to inhibit AAK1, we suggest it could be trialled, using an appropriate patient population with 2019-nCoV acute respiratory disease, to reduce both the viral entry and the inflammation in patients, using endpoints such as the MuLBSTA score, an early warning model for predicting mortality in viral pneumonia. 7 JS is editor-in-chief of Oncogene.…”

Identification and Optimization of 4‐Anilinoquinolines as Inhibitors of Cyclin G Associated Kinase

“…Inhibitors of the NAK family kinases have a high likelihood of showing low selectivity across the family based on high identity of their primary amino acid sequence and the presence of structural features which are distinct from those observed in other kinases. Specifically, the four NAK family kinases have a large α-helical insert positioned C -terminal to the activation segment, in proximity to the ATP binding site [34] . Notably, GAK does have one fewer amino acid residue in the hinge region, which distinguishes it from the other NAK family kinases.…”

“…Final assay volume for each data point was 5 μL, and final DMSO concentration was 1%. The kinase domain proteins were expressed in E. coli as a fusion with a C-terminal AVI tag (vector pNIC-Bio3, NCBI reference JN792439) which was biotinylated by co-expressed BirA, and purified using the same methods as used previously [34] . After setting up the assay plate it was incubated at room temperature for 1.5 hours and then read using a TR-FRET proto Residue ranges were AAK1: 31-396, BMP2K: 38-345, GAK: 12-347, STK16: 13-305col on a PheraStarFS plate reader (BMG Labtech).…”

Structural Basis for the Inhibition of Cyclin G‐Associated Kinase by Gefitinib

“…GAK is reportedly involved in the hepatitis C virus life cycle, and thus an inhibitor of GAK can be considered as an anti‐hepatitis C agent . Selective inhibitors for the NAK family kinases to inhibit the ATP binding site have been explored . Our results suggest that gefitinib binding at the second site is a selective interaction for both GAK and gefitinib.…”