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Selective covalent-allosteric tools to dissect Akt2

Quambusch, L.;D\'Angelo, G.;Kirschner, T.;Beerbaum, M.;Depta, L.;Schnecke, F.;Niggenaber, J.;Brandherm, S.;Weisner, J.;Mueller, M.;Dehmelt, L.;Rauh, D.

2026-06-19 Cancer Biology
10.64898/2026.06.15.732341 bioRxiv
Show abstract

The protein kinase Akt and its isoforms play a crucial role in various diseases. Unique functions of the individual isoforms (Akt1, Akt2, Akt3) might be essential for survival in malignancies. Particularly for Akt2, it was reported that a knock-out led to diabetic phenotype and might be correlated with clinically adverse hyperglycemic effects observed in pan Akt-treatment. Enduring failure of Akt inhibitors in the clinic indicates the necessity for a thorough understanding of the underlying biology, preferably by using highly isoform-selective small molecules. Here we report the structure-guided development of Akt2-selective covalent-allosteric probe molecules, that can be successfully modified within a complex environment using biorthogonal chemistry. Thus, enabling first Akt2-specific pull-down studies and the use in functional studies, such as selective fluorescent labeling in cellular systems. These chemical probes expand our toolbox to dissect the critical questions of Akt2s function in health and disease, thereby paving the way for novel therapeutic strategies based on thorough mechanistic insights.

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