Phosphorylation-Dependent Inhibition of Akt1
| Autor: | Xuguang Liu, Shawn S.-C. Li, McShane McKenna, Patrick O'Donoghue, Nileeka Balasuriya |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
inorganic chemicals lcsh:QH426-470 AKT1 Genetic code expansion environment and public health Article 03 medical and health sciences phosphoinositide dependent kinase 1 Protein kinase B In vivo Genetics biochemistry tRNASep Phosphoinositide dependent kinase 1 Genetics (clinical) chemistry.chemical_classification biology Chemistry TRNA sep Enzyme assay 3. Good health Pleckstrin homology domain phosphoseryl-tRNA synthetase Phosphoseryl-tRNA synthetase enzymes and coenzymes (carbohydrates) lcsh:Genetics 030104 developmental biology Enzyme genetic code expansion Protein kinase domain Biochemistry embryonic structures biology.protein Phosphorylation bacteria protein kinase B Phosphoinositide-dependent kinase-1 |
| Zdroj: | Genes, Vol 9, Iss 9, p 450 (2018) Genes Volume 9 Issue 9 Paediatrics Publications |
| ISSN: | 2073-4425 |
| Popis: | Protein kinase B (Akt1) is a proto-oncogene that is overactive in most cancers. Akt1 activation requires phosphorylation at Thr308 phosphorylation at Ser473 further enhances catalytic activity. Akt1 activity is also regulated via interactions between the kinase domain and the N-terminal auto-inhibitory pleckstrin homology (PH) domain. As it was previously difficult to produce Akt1 in site-specific phosphorylated forms, the contribution of each activating phosphorylation site to auto-inhibition was unknown. Using a combination of genetic code expansion and in vivo enzymatic phosphorylation, we produced Akt1 variants containing programmed phosphorylation to probe the interplay between Akt1 phosphorylation status and the auto-inhibitory function of the PH domain. Deletion of the PH domain increased the enzyme activity for all three phosphorylated Akt1 variants. For the doubly phosphorylated enzyme, deletion of the PH domain relieved auto-inhibition by 295-fold. We next found that phosphorylation at Ser473 provided resistance to chemical inhibition by Akti-1/2 inhibitor VIII. The Akti-1/2 inhibitor was most effective against pAkt1T308 and showed four-fold decreased potency with Akt1 variants phosphorylated at Ser473. The data highlight the need to design more potent Akt1 inhibitors that are effective against the doubly phosphorylated and most pathogenic form of Akt1. |
| Databáze: | OpenAIRE |
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