Article
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Preserved in Portico This version is not peer-reviewed
Phosphorylation-Dependent Inhibition of Akt1
Version 1
: Received: 20 July 2018 / Approved: 21 July 2018 / Online: 21 July 2018 (12:35:26 CEST)
A peer-reviewed article of this Preprint also exists.
Balasuriya, N.; McKenna, M.; Liu, X.; Li, S.S.C.; O’Donoghue, P. Phosphorylation-Dependent Inhibition of Akt1. Genes 2018, 9, 450. Balasuriya, N.; McKenna, M.; Liu, X.; Li, S.S.C.; O’Donoghue, P. Phosphorylation-Dependent Inhibition of Akt1. Genes 2018, 9, 450.
Abstract
Akt1 is a proto-oncogene that is over active 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-specifically 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 4-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.
Keywords
genetic code expansion, protein kinase B, phosphoinositide dependent kinase 1 (PDK1), phosphoseryl-tRNA synthetase (SepRS), tRNASep
Subject
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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