Umberto Maria Battisti, Chunxia Gao, Oscar Nilsson, Fady Akladios, Aleksei Lulla, Agnieszka Bogucka, Amalyn Nain-Perez, Liliana Håversen, Woonghee Kim, Jan Boren, Marko Hyvönen, Mathias Uhlen, Adil Mardinoglu, Morten Grøtli
ChemBioChem, accepted article 17th Oct (2022)
DOI: 0.1002/cbic.202200339
Pubmed: 36250581
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.