Multiple conformational states of the HPK1 kinase domain in complex with sunitinib reveal the structural changes accompanying HPK1 trans-regulation
| Autor: | Sergei Timofeevski, Sansana Sawasdikosol, Timothy Scott Fisher, Steven J. Burakoff, Robert Steven Kania, Ted William Johnson, Gallego Rebecca Anne, Ciarán N. Cronin, Eric Johnson, Michele McTigue, Michael F. Maestre |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
MAPK/ERK pathway T-Lymphocytes Serine threonine protein kinase Molecular Dynamics Simulation Protein Serine-Threonine Kinases Crystallography X-Ray Biochemistry 03 medical and health sciences Adenosine Triphosphate medicine Sunitinib Humans Phosphorylation Molecular Biology Binding Sites 030102 biochemistry & molecular biology Kinase Chemistry Cell Biology Recombinant Proteins Cell biology Protein Structure Tertiary 030104 developmental biology Protein kinase domain Accelerated Communications Mutagenesis Site-Directed Interleukin-2 Signal transduction Tyrosine kinase Dimerization medicine.drug |
| Zdroj: | J Biol Chem |
| ISSN: | 1083-351X |
| Popis: | Hematopoietic progenitor kinase 1 (HPK1 or MAP4K1) is a Ser/Thr kinase that operates via the c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) signaling pathways to dampen the T-cell response and antitumor immunity. Accordingly, selective HPK1 inhibition is considered a means to enhance antitumor immunity. Sunitinib, a multi-receptor tyrosine kinase (RTK) inhibitor approved for the management of gastrointestinal stromal tumors (GISTs), renal cell carcinoma (RCC), and pancreatic cancer, has been reported to inhibit HPK1 in vitro. In this report, we describe the crystal structures of the native HPK1 kinase domain in both nonphosphorylated and doubly phosphorylated states, in addition to a double phosphomimetic mutant (T165E,S171E), each complexed with sunitinib at 2.17–3.00-Å resolutions. The native nonphosphorylated cocrystal structure revealed an inactive dimer in which the activation loop of each monomer partially occupies the ATP- and substrate-binding sites of the partner monomer. In contrast, the structure of the protein with a doubly phosphorylated activation loop exhibited an active kinase conformation with a greatly reduced monomer–monomer interface. Conversely, the phosphomimetic mutant cocrystal structure disclosed an alternative arrangement in which the activation loops are in an extended domain-swapped configuration. These structural results indicate that HPK1 is a highly dynamic kinase that undergoes trans-regulation via dimer formation and extensive intramolecular and intermolecular remodeling of the activation segment. |
| Databáze: | OpenAIRE |
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