ATP-dependent phosphorylation of α-substituted carboxylic acids catalyzed by pyruvate kinase

Ash, David E.; Goodhart, Paula J.; Reed, George H.

doi:10.1016/0003-9861(84)90043-2

Cited by 31 publications

(22 citation statements)

References 28 publications

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“…Additionally, no enzyme capable of phosphorylating l -lactate to 2-PL has been identified in F 420 -producing organisms, despite considerable investigation 22 . 2-PL has been little studied as a metabolite and is only known to occur as a by-product of pyruvate kinase activity 27 . 2-PL has not been implicated as a substrate in any metabolic pathway outside the proposed role in F 420 biosynthesis; rather, it has been shown in vitro to inhibit several enzymes involved in glycolysis and amino acid biosynthesis 28–30 .…”

Section: Resultsmentioning

confidence: 99%

A revised biosynthetic pathway for the cofactor F420 in prokaryotes

et al. 2019

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Cofactor F 420 plays critical roles in primary and secondary metabolism in a range of bacteria and archaea as a low-potential hydride transfer agent. It mediates a variety of important redox transformations involved in bacterial persistence, antibiotic biosynthesis, pro-drug activation and methanogenesis. However, the biosynthetic pathway for F 420 has not been fully elucidated: neither the enzyme that generates the putative intermediate 2-phospho- l -lactate, nor the function of the FMN-binding C-terminal domain of the γ-glutamyl ligase (FbiB) in bacteria are known. Here we present the structure of the guanylyltransferase FbiD and show that, along with its archaeal homolog CofC, it accepts phosphoenolpyruvate, rather than 2-phospho- l -lactate, as the substrate, leading to the formation of the previously uncharacterized intermediate dehydro-F 420 -0. The C-terminal domain of FbiB then utilizes FMNH 2 to reduce dehydro-F 420 -0, which produces mature F 420 species when combined with the γ-glutamyl ligase activity of the N-terminal domain. These new insights have allowed the heterologous production of F 420 from a recombinant F 420 biosynthetic pathway in Escherichia coli .

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Section: Resultsmentioning

confidence: 99%

A revised biosynthetic pathway for the cofactor F420 in prokaryotes

et al. 2019

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“…Previous in vitro studies had suggested that 2-P-lactate can be produced when pyruvate kinase acts on L-lactate, an end product of glycolysis 20 . Indeed, incubation of recombinant pyruvate kinase M2 with L-lactate and ATP led to the formation of 2-P-L-lactate, albeit at a very slow rate (for example, 1.3 nmol min −1 mg −1 at 5 mM L-lactate) and with an estimated K m of more than 30 mM (compared to the forward reaction with a K m of 142 μM and a V max of 100 μmol min −1 mg −1 ; Fig.…”

Section: Increase Of 2-p-l-lactate and Decrease Of Fructose 26-bpmentioning

confidence: 99%

A conserved phosphatase destroys toxic glycolytic side products in mammals and yeast

et al. 2016

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Metabolic enzymes are very specific. However, most of them show weak side activities toward compounds that are structurally related to their physiological substrates, thereby producing side products that may be toxic. In some cases, 'metabolite repair enzymes' eliminating side products have been identified. We show that mammalian glyceraldehyde 3-phosphate dehydrogenase and pyruvate kinase, two core glycolytic enzymes, produce 4-phosphoerythronate and 2-phospho-L-lactate, respectively. 4-Phosphoerythronate strongly inhibits an enzyme of the pentose phosphate pathway, whereas 2-phospho-L-lactate inhibits the enzyme producing the glycolytic activator fructose 2,6-bisphosphate. We discovered that a single, widely conserved enzyme, known as phosphoglycolate phosphatase (PGP) in mammals, dephosphorylates both 4-phosphoerythronate and 2-phospho-L-lactate, thereby preventing a block in the pentose phosphate pathway and glycolysis. Its yeast ortholog, Pho13, similarly dephosphorylates 4-phosphoerythronate and 2-phosphoglycolate, a side product of pyruvate kinase. Our work illustrates how metabolite repair enzymes can make up for the limited specificity of metabolic enzymes and permit high flux in central metabolic pathways.

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“…Two kinases are known to potentially phosphorylate a hydroxyl group present on carbon 2 of a carboxylic acid. One is pyruvate kinase, which is known to display side activities that lead to the phosphorylation of lactate and glycolate [44]. Another one is glycerate kinase, which converts glycerate to 2-P-glycerate in vertebrates and in bacteria [45].…”

Section: Discussionmentioning

confidence: 99%

Phosphoglycolate has profound metabolic effects but most likely no role in a metabolic DNA response in cancer cell lines

Gerin

Bury

Baldin

et al. 2019

Biochemical Journal

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Repair of a certain type of oxidative DNA damage leads to the release of phosphoglycolate, which is an inhibitor of triose phosphate isomerase and is predicted to indirectly inhibit phosphoglycerate mutase activity. Thus, we hypothesized that phosphoglycolate might play a role in a metabolic DNA damage response. Here, we determined how phosphoglycolate is formed in cells, elucidated its effects on cellular metabolism and tested whether DNA damage repair might release sufficient phosphoglycolate to provoke metabolic effects. Phosphoglycolate concentrations were below 5 µM in wild-type U2OS and HCT116 cells and remained unchanged when we inactivated phosphoglycolate phosphatase (PGP), the enzyme that is believed to dephosphorylate phosphoglycolate. Treatment of PGP knockout cell lines with glycolate caused an up to 500-fold increase in phosphoglycolate concentrations, which resulted largely from a side activity of pyruvate kinase. This increase was much higher than in glycolate-treated wild-type cells and was accompanied by metabolite changes consistent with an inhibition of phosphoglycerate mutase, most likely due to the removal of the priming phosphorylation of this enzyme. Surprisingly, we found that phosphoglycolate also inhibits succinate dehydrogenase with a Ki value of <10 µM. Thus, phosphoglycolate can lead to profound metabolic disturbances. In contrast, phosphoglycolate concentrations were not significantly changed when we treated PGP knockout cells with Bleomycin or ionizing radiation, which are known to lead to the release of phosphoglycolate by causing DNA damage. Thus, phosphoglycolate concentrations due to DNA damage are too low to cause major metabolic changes in HCT116 and U2OS cells.

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ATP-dependent phosphorylation of α-substituted carboxylic acids catalyzed by pyruvate kinase

Cited by 31 publications

References 28 publications

A revised biosynthetic pathway for the cofactor F420 in prokaryotes

A revised biosynthetic pathway for the cofactor F420 in prokaryotes

A conserved phosphatase destroys toxic glycolytic side products in mammals and yeast

Phosphoglycolate has profound metabolic effects but most likely no role in a metabolic DNA response in cancer cell lines

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