| Affiliation: | 1. Department of Chemistry and Molecular Biology, University of Gothenburg, 412 96 Gothenburg, Sweden;2. Department of Chemistry and Molecular Biology, University of Gothenburg, 412 96 Gothenburg, Sweden These authors contributed equally to this work.;3. Department of Biochemistry, University of Cambridge, Cambridge, CB2 1GA UK;4. Department of Molecular and Clinical Medicine, University of Gothenburg and Sahlgrenska University Hospital, 413 45 Gothenburg, Sweden;5. Science for Life Laboratory, KTH-Royal Institute of Technology, 171 21 Stockholm, Sweden |
| Abstract: | Enzymes are effective biological catalysts that accelerate almost all metabolic reactions in living organisms. Synthetic modulators of enzymes are useful tools for the study of enzymatic reactions and can provide starting points for the design of new drugs. Here, we report on the discovery of a class of biologically active compounds that covalently modifies lysine residues in human liver pyruvate kinase (PKL), leading to allosteric activation of the enzyme (EC50=0.29 μM). Surprisingly, the allosteric activation control point resides on the lysine residue K282 present in the catalytic site of PKL. These findings were confirmed by structural data, MS/MS experiments, and molecular modelling studies. Altogether, our study provides a molecular basis for the activation mechanism and establishes a framework for further development of human liver pyruvate kinase covalent activators. |
