New views on an old enzyme: allosteric regulation and evolution of archaeal pyruvate kinases. (26th April 2019)

• Record Type: Journal Article
• Authors: Johnsen, Ulrike; Reinhardt, Andreas; Landan, Giddy; Tria, Fernando D. K.; Turner, Jonathan M.; Davies, Christopher; Schönheit, Peter
• Is Part Of: FEBS journal. Volume 286:Number 13(2019)
• Page Start: 2471
• Page End: 2489
• Publication Date: 2019-04-26
• Subjects: 3‐phosphoglycerate -- allosteric regulation -- archaea -- evolution -- pyruvate kinase; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pathology, Molecular -- Periodicals; 572
• DOI: 10.1111/febs.14837
• Languages: English
• ISSNs: 1742-464X

Abstract

Abstract : Pyruvate kinases (PKs) synthesize ATP as the final step of glycolysis in the three domains of life. PKs from most bacteria and eukarya are allosteric enzymes that are activated by sugar phosphates; for example, the feed‐forward regulator fructose‐1, 6‐bisphosphate, or AMP as a sensor of energy charge. Archaea utilize unusual glycolytic pathways, but the allosteric properties of PKs from these species are largely unknown. Here, we present an analysis of 24 PKs from most archaeal clades with respect to allosteric properties, together with phylogenetic analyses constructed using a novel mode of rooting protein trees. We find that PKs from many Thermoproteales, an order of crenarchaeota, are allosterically activated by 3‐phosphoglycerate (3PG). We also identify five conserved amino acids that form the binding pocket for 3PG. 3PG is generated via an irreversible reaction in the modified glycolytic pathway of these archaea and therefore functions as a feed‐forward regulator. We also show that PKs from hyperthermophilic Methanococcales, an order of euryarchaeota, are activated by AMP. Phylogenetic analyses indicate that 3PG‐activated PKs form an evolutionary lineage that is distinct from that of sugar‐phosphate activated PKs, and that sugar phosphate‐activated PKs originated as AMP‐regulated PKs in hyperthermophilic Methanococcales. Since the phospho group of sugar phosphates and 3PG overlap in the allosteric site, our data indicate that the allostery in PKs first started from a progenitor phosphate‐binding site that evolved in two spatially distinct directions: one direction generated the canonical site that responds to sugar phosphates and the other gave rise to the 3PG site present in Thermoproteales. Overall, our data suggest an intimate connection between the allosteric properties and evolution of PKs. Abstract : Pyruvate kinases (PKs) are allosteric enzymes that catalyze the final step of glycolysis. In most species, PKs are activated by sugar phosphates such as fructose bisphosphate or AMP. In this study, Peter Schönheit and colleagues report that PKs of the archaeal group Thermoproteales are activated by a novel effector, 3‐phosphoglycerate (3PG). They also identify conserved amino acids that form a 3PG‐binding pocket. The authors propose that allostery in this enzyme evolved from a simple phosphate‐binding site that expanded in two structurally distinct directions: one resulting in canonical activation by sugar phosphates and the other by 3PG.