Serendipitous Identification of a Covalent Activator of Liver Pyruvate Kinase

• 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. 

Ämnesord

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)

Nyckelord

covalent activators

enzymes

ligand design

PKL

structure-activity relationships

benzo[d]isothiazole 1

1 dioxide

chloro saccharin

enzyme inhibitor

isoenzyme

lysine

phosphoenolpyruvate

protein PKL

protein PKM1

protein PKM2

protein PKR

pyruvate kinase

saccharin

unclassified drug

allosterism

aqueous solution

Article

biocatalysis

biochemical analysis

computer model

controlled study

EC50

enzyme activation

enzyme active site

enzyme modification

enzyme structure

erythrocyte

glycolysis

human

in vitro study

liver

mass spectrometry

molecular docking

molecular dynamics

molecular stability

protein analysis

protein expression

RNA splicing

synthesis

covalent activators

enzymes

ligand design

PKL

structure-activity

relationships

inhibitors

isoenzymes

discovery

binding

protein

site

Biochemistry & Molecular Biology

Pharmacology & Pharmacy

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