Homology modeling

The H. sapiens HUNK sequence was retrieved from Uniprot ({"type":"entrez-protein","attrs":{"text":"P57058","term_id":"9973393","term_text":"P57058"}}P57058) and used in a protein-protein blast against the Protein Data Bank (PDB). The top scoring PDB structures were retrieved. In addition all protein kinase structure resolved with staurosporine was retrieved from the Protein Data Bank (PDB). A protein sequence alignment was performed between HUNK and the retrieved protein kinases using MUSCLE [29]. Manual edits were performed using Discovery Studio (Dassault Systèmes BIOVIA, Discovery Studio, Release 2017, San Diego: Dassault Systèmes, 2018). Homology models bound with staurosporine were generated from two template structures (PDB entries: 1NVR and 1WVY) using modeler 9v19 [30]. Amino acids are numbered with the initiating methionine set to 1. The models were subjected to quality analysis using the PDBsum generator (http://www.ebi.ac.uk/pdbsum) [31]. The models were prepared for analysis using the protein preparation wizard in which protonation states were assigned followed by an energy minimization to relieve unfavorable constraints (Schrödinger Release 2017-3 Schrödinger, LLC, New York, NY, 2017).