TY - JOUR T1 - Functional cross-talk between allosteric effects of activating and inhibiting ligands underlies PKM2 regulation JF - bioRxiv DO - 10.1101/378133 SP - 378133 AU - Jamie A. Macpherson AU - Alina Theisen AU - Laura Masino AU - Louise Fets AU - Paul C. Driscoll AU - Vesela Encheva AU - Ambrosius P. Snijders AU - Stephen R. Martin AU - Jens Kleinjung AU - Perdita E. Barran AU - Franca Fraternali AU - Dimitrios Anastasiou Y1 - 2018/01/01 UR - http://biorxiv.org/content/early/2018/07/26/378133.abstract N2 - Allosteric regulation is central to the role of the glycolytic enzyme pyruvate kinase M2 (PKM2) in cellular metabolism. Multiple activating and inhibitory allosteric ligands regulate PKM2 activity by controlling the equilibrium between high activity tetramers and low activity dimers and monomers. However, it remains elusive how allosteric inputs upon simultaneous binding of different ligands are integrated to regulate PKM2 activity. Here, we show that, in the presence of the allosteric inhibitor L-phenylalanine (Phe), the activator fructose 1,6-bisphosphate (FBP) can induce PKM2 tetramerisation, but fails to maximally increase enzymatic activity. Guided by a new computational framework we developed to identify residues that mediate FBP-induced allostery, we generated two PKM2 mutants, A327S and C358A, in which activation by FBP remains intact but cannot be attenuated by Phe. Our findings demonstrate a role for residues involved in FBP-induced allostery in enabling the integration of allosteric input from Phe and reveal a mechanism that underlies the co-ordinate regulation of PKM2 activity by multiple allosteric ligands. ER -