Elucidation of a Self-Sustaining Cycle in <i>Escherichia coli</i> <scp>l</scp>-Serine Biosynthesis That Results in the Conservation of the Coenzyme, NAD<sup>+</sup>

Grant, Gregory A.

doi:10.1021/acs.biochem.8b00074

Cited by 15 publications

(26 citation statements)

References 35 publications

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“…However, given our previous observation that Ser3 and Ser33 reduce αKG to the D enantiomer of this dicarboxylic acid [38], this is most likely also the product of the 3PGA-αKG transhydrogenase activity. While this work was ongoing, other investigators independently showed that the Pseudomonas and E. coli phosphoglycerate dehydrogenases (SerA) also actually act as 3PGA-αKG transhydrogenases that produce D-2HG [52,53], perfectly corroborating our results with the yeast enzymes. Only relatively few examples of transhydrogenases are known so far.…”

Section: Nad(p)hx Repair Deficiency Leads To Discrete Changes In Genesupporting

confidence: 84%

NAD(P)HX repair deficiency causes central metabolic perturbations in yeast and human cells

Becker-Kettern

Paczia

Conrotte

et al. 2018

Preprint

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reading frame; PSAT1, phosphoserine transaminase 1. Keywords-metabolite repair, NAD(P)HX dehydratase, NAD(P)HX epimerase, Saccharomyces cerevisiae, inborn errors of metabolism.Conflict of interest-The authors declare that they have no conflicts of interest with the contents of this article.. CC-BY-NC-ND 4.0 International license It is made available under a (which was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint . http://dx.doi.org/10.1101/302257 doi: bioRxiv preprint first posted online Apr. 16, 2018; 2 ABSTRACT NADHX and NADPHX are hydrated and redox inactive forms of the NADH and NADPH cofactors, known to inhibit several dehydrogenases in vitro. A metabolite repair system that is conserved in all domains of life and that comprises the two enzymes NAD(P)HX dehydratase and NAD(P)HX epimerase, allows reconversion of both the S-and R-epimers of NADHX and NADPHX to the normal cofactors. An inherited deficiency in this system has recently been shown to cause severe neurometabolic disease in children. Although evidence for the presence of NAD(P)HX has been obtained in plant and human cells, little is known about the mechanism of formation of these derivatives in vivo and their potential effects on cell metabolism. Here, we show that NAD(P)HX dehydratase deficiency in yeast leads to an important, temperature-dependent NADHX accumulation in quiescent cells with a concomitant depletion of intracellular NAD + and serine pools. We demonstrate that NADHX potently inhibits the first step of the serine synthesis pathway in yeast. Human cells deficient in the NAD(P)HX dehydratase also accumulated NADHX and showed decreased viability. In addition, those cells consumed more glucose and produced more lactate, potentially indicating impaired mitochondrial function. Our results provide first insights into how NADHX accumulation affects cellular functions and pave the way for a better understanding of the mechanism(s) underlying the rapid and severe neurodegeneration leading to early death in NADHX repair deficient children.

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Section: Nad(p)hx Repair Deficiency Leads To Discrete Changes In Genesupporting

confidence: 84%

NAD(P)HX repair deficiency causes central metabolic perturbations in yeast and human cells

Becker-Kettern

Paczia

Conrotte

et al. 2018

Preprint

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“…However, given our previous observation that Ser3 and Ser33 reduce αKG to the D enantiomer of this dicarboxylic acid , this is most likely also the product of the 3PGA‐αKG transhydrogenase activity. While this work was ongoing, other investigators independently showed that the Pseudomonas and E. coli phosphoglycerate dehydrogenases (SerA) also actually act as 3PGA‐αKG transhydrogenases that produce D‐2HG , perfectly corroborating our results with the yeast enzymes. Only relatively few examples of transhydrogenases are known so far.…”

Section: Discussionsupporting

confidence: 85%

NAD(P)HX repair deficiency causes central metabolic perturbations in yeast and human cells

et al. 2018

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NADHX and NADPHX are hydrated and redox inactive forms of the NADH and NADPH cofactors, known to inhibit several dehydrogenases in vitro. A metabolite repair system that is conserved in all domains of life and that comprises the two enzymes NAD(P)HX dehydratase and NAD(P)HX epimerase, allows reconversion of both the S- and R-epimers of NADHX and NADPHX to the normal cofactors. An inherited deficiency in this system has recently been shown to cause severe neurometabolic disease in children. Although evidence for the presence of NAD(P)HX has been obtained in plant and human cells, little is known about the mechanism of formation of these derivatives in vivo and their potential effects on cell metabolism. Here, we show that NAD(P)HX dehydratase deficiency in yeast leads to an important, temperature-dependent NADHX accumulation in quiescent cells with a concomitant depletion of intracellular NAD and serine pools. We demonstrate that NADHX potently inhibits the first step of the serine synthesis pathway in yeast. Human cells deficient in the NAD(P)HX dehydratase also accumulated NADHX and showed decreased viability. In addition, those cells consumed more glucose and produced more lactate, potentially indicating impaired mitochondrial function. Our results provide first insights into how NADHX accumulation affects cellular functions and pave the way for a better understanding of the mechanism(s) underlying the rapid and severe neurodegeneration leading to early death in NADHX repair-deficient children.

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“…Values are per tetramer .b From Zhao and Winkler [29] .c ND, not determined; DNI, does not inhibit; DNR, does not react; NDS, not detectable spectrophotometrically at 340 nm .d Value varies slightly from different groups. Note that in all cases, all of the kinetic parameters can vary depending on the buffer used .e Exhibits substantial substrate inhibition in phosphate buffer, pH 7.0 .f Unpublished .g Estimated from Achouri et al (1997) .h From Grant (2018) .…”

Section: Regulation Of Catalytic Activitymentioning

confidence: 99%

“…Zhang et al (Zhang et al, 2017) have shown in Pseudomonas species, that the coupling of this reaction with d-2-hydroxyglutarate dehydrogenase can serve to drive l -serine synthesis. Grant (2018) has shown that in E. coli , there is a process that conserves coenzyme in the production of l -serine by utilizing an intrinsic cycle of NAD + /NADH interconversion coupled with the conversion of αKG to αHG. Interestingly, this cycle can be maintained in vitro by production of αKG by the second enzyme in the pathway, PSAT, and does not require any additional enzymes (Figure 11).…”

Section: Alternate Substratesmentioning

confidence: 99%

“…From a structural and mechanistic point of view, the most studied PGDH is that from E. coli (Pizer, 1963; Pizer and Potochny, 1964; Rosenbloom et al, 1968; Sugimoto and Pizer, 1968a,b; Winicov and Pizer, 1974; Dubrow and Pizer, 1977a,b; McKitrick and Pizer, 1980; Tobey and Grant, 1986; Schuller et al, 1995; Al-Rabiee et al, 1996a,b; Grant et al, 1996, 1999a,b, 2000a,b, 2001a,b, 2002, 2003, 2004, 2005; Zhao and Winkler, 1996; Grant and Xu, 1998; Bell et al, 2002, 2004; Grant, 2004, 2011, 2012, 2018; Thompson et al, 2005; Dey et al, 2007; Burton et al, 2008, 2009a), followed by that from M. tuberculosis (Grant et al, 1999c; Dey et al, 2005a,b, 2008; Burton et al, 2007, 2009b; Xu and Grant, 2014; Xu et al, 2015). There are also reports from various animal tissues (Pizer, 1964; Walsh and Sallach, 1965; Cheung et al, 1969; Pizer and Sugimoto, 1971; Grant and Bradshaw, 1978; Grant et al, 1978; Lund et al, 1986; Fell and Snell, 1988; Achouri et al, 1997), other eukaryotes (Ulane and Ogur, 1972; Ali et al, 2004; Singh et al, 2014), other bacteria (Umbarger and Umbarger, 1962; Umbarger et al, 1963; Saski and Pizer, 1975; Peters-Wendisch et al, 2002, 2005), and plants (Hanford and Davies, 1958; Cheung et al, 1968; Slaughter and Davies, 1968a,b; Rosenblum and Sallach, 1970).…”

Section: Introductionmentioning

confidence: 99%

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D-3-Phosphoglycerate Dehydrogenase

Grant

2018

Front. Mol. Biosci.

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l-Serine is the immediate precursor of d-serine, a major agonist of the N-methyl-d-aspartate (NMDA) receptor. l-Serine is a pivotal amino acid since it serves as a precursor to a large number of essential metabolites besides d-serine. In all non-photosynthetic organisms, including mammals, a major source of l-serine is the phosphorylated pathway of l-serine biosynthesis. The pathway consists of three enzymes, d-3-phosphoglycerate dehydrogenase (PGDH), phosphoserine amino transferase (PSAT), and l-phosphoserine phosphatase (PSP). PGDH catalyzes the first step in the pathway by converting d-3-phosphoglycerate (PGA), an intermediate in glycolysis, to phosphohydroxypyruvate (PHP) concomitant with the reduction of NAD+. In some, but not all organisms, the catalytic activity of PGDH can be regulated by feedback inhibition by l-serine. Three types of PGDH can be distinguished based on their domain structure. Type III PGDHs contain only a nucleotide binding and substrate binding domain. Type II PGDHs contain an additional regulatory domain (ACT domain), and Type I PGDHs contain a fourth domain, termed the ASB domain. There is no consistent pattern of domain content that correlates with organism type, and even when additional domains are present, they are not always functional. PGDH deficiency results in metabolic defects of the nervous system whose systems range from microcephaly at birth, seizures, and psychomotor retardation. Although deficiency of any of the pathway enzymes have similar outcomes, PGDH deficiency is predominant. Dietary or intravenous supplementation with l-serine is effective in controlling seizures but has little effect on psychomotor development. An increase in PGDH levels, due to overexpression, is also associated with a wide array of cancers. In culture, PGDH is required for tumor cell proliferation, but extracellular l-serine is not able to support cell proliferation. This has led to the hypothesis that the pathway is performing some function related to tumor growth other than supplying l-serine. The most well-studied PGDHs are bacterial, primarily from Escherichia coli and Mycobacterium tuberculosis, perhaps because they have been of most interest mechanistically. However, the relatively recent association of PGDH with neuronal defects and human cancers has provoked renewed interest in human PGDH.

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Elucidation of a Self-Sustaining Cycle in Escherichia coli l-Serine Biosynthesis That Results in the Conservation of the Coenzyme, NAD⁺

Cited by 15 publications

References 35 publications

NAD(P)HX repair deficiency causes central metabolic perturbations in yeast and human cells

NAD(P)HX repair deficiency causes central metabolic perturbations in yeast and human cells

NAD(P)HX repair deficiency causes central metabolic perturbations in yeast and human cells

D-3-Phosphoglycerate Dehydrogenase

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Elucidation of a Self-Sustaining Cycle in Escherichia coli l-Serine Biosynthesis That Results in the Conservation of the Coenzyme, NAD+

Cited by 15 publications

References 35 publications

NAD(P)HX repair deficiency causes central metabolic perturbations in yeast and human cells

NAD(P)HX repair deficiency causes central metabolic perturbations in yeast and human cells

NAD(P)HX repair deficiency causes central metabolic perturbations in yeast and human cells

D-3-Phosphoglycerate Dehydrogenase

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Elucidation of a Self-Sustaining Cycle in Escherichia coli l-Serine Biosynthesis That Results in the Conservation of the Coenzyme, NAD⁺