Purification and comparative properties of the glycoprotein nicotinamide adenine dinucleotide glycohydrolase from rat liver microsomal and plasma membranes

DiAugustine, R P; Abe, Tetsuaki; Voytek, Peter; Eckberg, Dolores

doi:10.1016/0005-2744(78)90142-0

Cited by 13 publications

(5 citation statements)

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“…Other studies on the rat liver enzyme failed to demonstrate a clear-cut difference between the NAD glycohydrolase of microsomes and that of plasma membrane preparations. Both enzymes have nearly the same half-life (19), give a single continuous precipitation line in Ouchterlony double-diffusion immunochemical tests (18), and show similar biochemical characteristics (24). Such experimental data are consistent with NAD glycohydrolase being a constituent of a single membrane entity in the liver.…”

Section: Nad Glycohydrolase Activity In Sinusoidal Cells Obtained By Counterflow Elutriationsupporting

confidence: 71%

“…The presence of NAD glycohydrolase in the endoplasmic reticulum implies a rapid turnover of the cytosolic pyridine coenzymes, as recalled above, if no compartmental hindrance to their breakdown is present. In addition, except for minor differences in carbohydrate composition, comparative studies on the plasmalemmal and microsomal NAD glycohydrolase from rat liver did not reveal noticeable differences in their biochemical (24) and immunological (18) properties, and in their half-lives (19). Finally, there is at present no clear-cut example of an enzyme being a constituent of the endoplasmic reticulum and of plasma membranes in liver cells (11,47).…”

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confidence: 99%

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Analytical study of microsomes and isolated subcellular membranes from rat liver. IX. Nicotinamide adenine dinucleotide glycohydrolase: a plasma membrane enzyme prominently found in Kupffer cells.

Amar-Costesec¹,

Pradofigueroa²,

Beaufay³

et al. 1985

The Journal of Cell Biology

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The distribution of nicotinamide adenine dinucleotide (NAD) glycohydrolase in rat liver was investigated by subcellular fractionation and by isolation of hepatocytes and sinusoidal cells. The behavior of NAD glycohydrolase in subcellular fractionation was peculiar because, although the enzyme was mainly microsomal, plasma membrane preparations contained distinctly more NAD glycohydrolase than could be accounted for by their content in elements derived from the endoplasmic reticulum or the Golgi complex identified by glucose-6-phosphatase and galactosyltransferase, respectively. When microsomal and plasmalemmal preparations were brought to equilibrium in a linear-density gradient, NAD glycohydrolase differed from these enzymes and behaved like 5'-nucleotidase and alkaline phosphodiesterase I. NAD glycohydrolase was markedly displaced towards higher densities after treatment with digitonin. This behavior in density-gradient centrifugation strongly suggests that NAD glycohydrolase is an exclusive enzyme of the plasma membrane. NAD glycohydrolase differed clearly from other plasmalemmal enzymes when the liver was fractionated into hepatocytes and sinusoidal cells; its specific activity was considerably greater in sinusoidal cell than in hepatocyte preparations. Further subfractionation of sinusoidal cell preparations into endothelial and Kupffer cells by counterflow elutriation showed that NAD glycohydrolase is more active in Kupffer cells. We estimate that the specific activity of NAD glycohydrolase activity is at least 65-fold higher at the periphery of Kupffer cells than at the periphery of hepatocytes. As the enzyme shows no structure-linked latency and is an exclusive constituent of the plasma membranes, we conclude that it is an ectoenzyme that cannot lead to a rapid turnover of the cytosolic pyridine nucleotides.The nicotinamide adenine dinucleotide (NAD) ~ glycohydrolase reaction appears barely consistent with the metabolic role Abbreviations used in this paper: NAD, nicotinamide adenine dinucleotide; NPC~ and NPC2, nonparenchymal cells; PC, parenchymal cells.of the pyridine nucleotides, particulady in the liver. Indeed, the high activity ofNAD glycohydrolase in hepatocytes should result in a rapid turnover of the pyridine coenzymes through the cycle proposed by Gholson (26). The resynthesis of NAD from nicotinamide requires three ATP molecules and expends

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Section: Nad Glycohydrolase Activity In Sinusoidal Cells Obtained By Counterflow Elutriationsupporting

confidence: 71%

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confidence: 99%

Analytical study of microsomes and isolated subcellular membranes from rat liver. IX. Nicotinamide adenine dinucleotide glycohydrolase: a plasma membrane enzyme prominently found in Kupffer cells.

Amar-Costesec¹,

Pradofigueroa²,

Beaufay³

et al. 1985

The Journal of Cell Biology

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“…From results derived in a long series of experiments utilizing NAD + labelled in adenine and nicotiinic moieties (Mandel and Amos, manuscript submitted for publication), designed to answer the question of whether any intact NAD+ can be transported into Nil cells, we are forced to conclude that no detectable NAD+ can be demonstrated to accumulate in Nil cells under a variety of experimental conditions. As do many other cells, Nil cells possess a highly active NAD +-glycohydrolase activity on the plasma membrane surface (ecto-NADase) which rapidly h:ydrolyses the nicotinamide-riboside bond releasing nicotinamide plus ADP-ribose from NAD' (Pekala and Anderson, 1978;DiAugustine et al, 1978;Artman and Seeley, 1979;Pekala et al, 1980). Upon addition of 10 pM NAD + to the cell monolayers, the ecto-NADase activity hydrolyses virtually all the exogenously added NAD + within 30 to 45 minutes (data not shown) and thus little intact NAD+ is left available for accumulation.…”

Section: Effect Of Nicotinamide Deprivation On Intracellularmentioning

confidence: 99%

Reactivation of NAD(H) biosynthetic pathway by exogenous NAD⁺ in nil cells severely depleted of NAD(H)

Mandel

Lively

Lombardi

et al. 1983

Journal Cellular Physiology

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The culture of Nil hamster fibroblasts in MEM lacking nicotinamide (NAm-MEM) leads to: (1) the rapid loss of intracellular total nicotinamide adenine dinucleotide (NAD(H)) content in these cells from a level of 150-200 pmoles/10(5) cells to less than 20 pmoles/10(5) cells; (2) the cessation of cell division and inhibition of DNA synthesis; and (3) a reduction of glucose consumption and lactic acid production. In most situations, following nicotinamide starvation, the restoration of intracellular NAD(H) follows rapidly the readdition of NAD+ (oxidized), nicotinamide mononucleotide (NMN), nicotinamide, or nicotinic acid. Resumption of cell division occurs after only a lag of about 24 hours. Nil cells subcultured for three consecutive times in the absence of nicotinamide (3(0) NAm- cells) exhibit different behavior. These severely starved cells are incapable of quickly restoring their intracellular NAD(H) content to normal levels when provided with any pyridine ring compound except NAD+. One-hour exposure of such cells to NAD+ allows utilization of nicotinamide to rapidly restore intracellular NAD(H). This short incubation with NAD+ does not result in any significant restoration of intracellular NAD(H) or lead to the accumulation of an intracellular pool of some precursor. This function of NAD+ as a stimulatory signal to the NAD(H)-biosynthetic pathway in severely starved Nil cells is a previously unreported role of NAD+, and does not require protein synthesis.

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“…Earlier studies have indicated that, e.g. in liver, NAD+ glycohydrolase is a constituent of endoplasmic reticulum, plasma membranes (Bock et al, 1971; Diaugustine et al, 1978), nuclear envelopes (Green & Dobrjansky, 1972;Fukushima et al, 1976;Tamulevicius et al, 1979) and secondary lysosomes (Mellors et al, 1975). The physiological role of this enzyme remains elusive; it was thought to control intracellular concentrations of NAD+ (Kaplan, 1966;Johnson, 1980), but its occurrence at high concentrations in certain tissues was difficult to reconcile with the known turnover rates of the coenzyme in vivo (Bock et al, 1968;Clark & Pinder, 1969;Bernofsky & Pankov, 1973).…”

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confidence: 99%

NAD+ glycohydrolase, an ecto-enzyme of calf spleen cells

Muller¹,

Muller²,

Schuber³

1983

Biochemical Journal

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By using a sensitive fluorimetric assay of NAD+ glycohydrolase (EC 3.2.2.6), we showed that calf spleen cells are able to hydrolyse 1,N6-etheno-NAD+ given in the medium. The observed rates of substrate hydrolysis and product accumulation in the medium are equivalent. Moreover, the splenocytes are able to cleave the nicotinamide-ribose bond of a water-soluble polymer of NAD+, and their NAD+ glycohydrolase activity is fully inhibited by a high-molecular-weight Blue Dextran. Selective permeation of the cellular membrane digitonin revealed an intracellular pool of NAD+ glycohydrolase, which accounts for 25% of the total activity. We conclude that NAD+ glycohydrolase associated with the splenocytes has the characteristics of an ecto-enzyme.

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Purification and comparative properties of the glycoprotein nicotinamide adenine dinucleotide glycohydrolase from rat liver microsomal and plasma membranes

Cited by 13 publications

References 38 publications

Analytical study of microsomes and isolated subcellular membranes from rat liver. IX. Nicotinamide adenine dinucleotide glycohydrolase: a plasma membrane enzyme prominently found in Kupffer cells.

Analytical study of microsomes and isolated subcellular membranes from rat liver. IX. Nicotinamide adenine dinucleotide glycohydrolase: a plasma membrane enzyme prominently found in Kupffer cells.

Reactivation of NAD(H) biosynthetic pathway by exogenous NAD⁺ in nil cells severely depleted of NAD(H)

NAD+ glycohydrolase, an ecto-enzyme of calf spleen cells

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Purification and comparative properties of the glycoprotein nicotinamide adenine dinucleotide glycohydrolase from rat liver microsomal and plasma membranes

Cited by 13 publications

References 38 publications

Analytical study of microsomes and isolated subcellular membranes from rat liver. IX. Nicotinamide adenine dinucleotide glycohydrolase: a plasma membrane enzyme prominently found in Kupffer cells.

Analytical study of microsomes and isolated subcellular membranes from rat liver. IX. Nicotinamide adenine dinucleotide glycohydrolase: a plasma membrane enzyme prominently found in Kupffer cells.

Reactivation of NAD(H) biosynthetic pathway by exogenous NAD+ in nil cells severely depleted of NAD(H)

NAD+ glycohydrolase, an ecto-enzyme of calf spleen cells

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Reactivation of NAD(H) biosynthetic pathway by exogenous NAD⁺ in nil cells severely depleted of NAD(H)