Functional cross-talk between allosteric effects of activating and inhibiting ligands underlies PKM2 regulation.
journal contributionposted on 09.01.2020 by Jamie A Macpherson, Alina Theisen, Laura Masino, Louise Fets, Paul C Driscoll, Vesela Encheva, Ambrosius P Snijders, Stephen R Martin, Jens Kleinjung, Perdita E Barran, Franca Fraternali, Dimitrios Anastasiou
Any type of content formally published in an academic journal, usually following a peer-review process.
Several enzymes can simultaneously interact with multiple intracellular metabolites, however, how the allosteric effects of distinct ligands are integrated to coordinately control enzymatic activity remains poorly understood. We addressed this question using, as a model system, the glycolytic enzyme pyruvate kinase M2 (PKM2). We show that the PKM2 activator fructose 1,6-bisphosphate (FBP) alone promotes tetramerisation and increases PKM2 activity, but addition of the inhibitor L-phenylalanine (Phe) prevents maximal activation of FBP-bound PKM2 tetramers. We developed a method, AlloHubMat, that uses eigenvalue decomposition of mutual information derived from molecular dynamics trajectories to identify residues that mediate FBP-induced allostery. Experimental mutagenesis of these residues identified PKM2 variants in which activation by FBP remains intact but cannot be attenuated by Phe. Our findings reveal residues involved in FBP-induced allostery that enable the integration of allosteric input from Phe and provide a paradigm for the coordinate regulation of enzymatic activity by simultaneous allosteric inputs.