PH domain-mediated autoinhibition and oncogenic activation of Akt
- Harvard Medical School, United States
- Dana-Farber Cancer Institute, United States
- The Ohio State University, United States
- Brigham and Women's Hospital, United States
Abstract
Akt is a Ser/Thr protein kinase that plays a central role in metabolism and cancer. Regulation of Akt's activity involves an autoinhibitory intramolecular interaction between its pleckstrin homology (PH) domain and its kinase domain that can be relieved by C-tail phosphorylation. PH domain mutant E17K Akt is a well-established oncogene. Previously, we reported that the conformation of autoinhibited Akt may be shifted by small molecule allosteric inhibitors limiting the mechanistic insights from existing X-ray structures that have relied on such compounds (Chu, Viennet, et al, 2020). Here we discover unexpectedly that a single mutation R86A Akt exhibits intensified autoinhibitory features with enhanced PH domain-kinase domain affinity. Structural and biochemical analysis uncovers the importance of a key interaction network involving Arg86, Glu17, and Tyr18 that controls Akt conformation and activity. Our studies also shed light on the molecular basis for E17K Akt activation as an oncogenic driver.
Data availability
Diffraction data have been deposited in PDB under the accession code: 7MYXSource data are uploaded as Zip files on the eLife website
Article and author information
Author details
Funding
National Cancer Institute (R01CA74305)
- Philip A Cole
National Cancer Institute (R35CA242461)
- Michael J Eck
National Cancer Institute (K22CA241105)
- Nam Chu
Claudia Adams Barr Program (Grant)
- Haribabu Arthanari
National Institutes of Health (EB002026)
- Haribabu Arthanari
Kwanjeong Education Foundation (Fellowship)
- Hwan Bae
National Institute of General Medical Sciences (GM124165)
- Michael J Eck
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2022, Bae et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
PH domain-mediated autoinhibition and oncogenic activation of Akt
eLife 11:e80148.
https://doi.org/10.7554/eLife.80148
Further reading
-
- Biochemistry and Chemical Biology
- Structural Biology and Molecular Biophysics
- Cell Biology
Our latest Special Issue brings together research on the allosteric regulation of kinase activity.
-
- Biochemistry and Chemical Biology
The protein ligase Connectase can be used to fuse proteins to small molecules, solid carriers, or other proteins. Compared to other protein ligases, it offers greater substrate specificity, higher catalytic efficiency, and catalyzes no side reactions. However, its reaction is reversible, resulting in only 50% fusion product from two equally abundant educts. Here, we present a simple method to reliably obtain 100% fusion product in 1:1 conjugation reactions. This method is efficient for protein-protein or protein-peptide fusions at the N- or C-termini. It enables the generation of defined and completely labeled antibody conjugates with one fusion partner on each chain. The reaction requires short incubation times with small amounts of enzyme and is effective even at low substrate concentrations and at low temperatures. With these characteristics, it presents a valuable new tool for bioengineering.
