Functional cross-talk between allosteric effects of activating and inhibiting ligands underlies PKM2 regulation

TY - JOUR

T1 - Functional cross-talk between allosteric effects of activating and inhibiting ligands underlies PKM2 regulation

AU - Macpherson, Jamie A.

AU - Theisen, Alina

AU - Masino, Laura

AU - Fets, Louise

AU - Driscoll, Paul C.

AU - Encheva, Vesela

AU - Snijders, Ambrosius P.

AU - Martin, Stephen R.

AU - Kleinjung, Jens

AU - Barran, Perdita E.

AU - Fraternali, Franca

AU - Anastasiou, Dimitrios

PY - 2019/7/2

Y1 - 2019/7/2

N2 - 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.

AB - 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.

KW - allostery

KW - computational biology

KW - enzymology

KW - molecular biophysics

KW - molecular dynamics

KW - native mass spectrometry

KW - none

KW - structural biology

KW - systems biology

UR - http://www.scopus.com/inward/record.url?scp=85070056233&partnerID=8YFLogxK

U2 - 10.7554/eLife.45068

DO - 10.7554/eLife.45068

M3 - Article

C2 - 31264961

AN - SCOPUS:85070056233

SN - 2050-084X

VL - 8

JO - eLife

JF - eLife

M1 - e45068

ER -