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A note on the ADP-ribose-protein linkages in rat-liver nuclei: a possible approach to assessing megavitamin therapy with niacin

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Summary

Nuclei isolated from rat liver were incubated with NAD⊕ whose two ribose moieties were respectively labeled with3H or14C. By enzymatic (phosphodiesterase) and/or chemical (hydroxylamine) attack on doubly labeled ADP-ribosylated nuclear proteins and identification of fragments released from nuclear residues, AMP was found after hydroxylaminolysis as well as iso-ADP-ribose after phosphodiesterase plus hydroxylamine, in the absence of detectable amounts of ribose-5-phosphate. This is taken to indicate the existence of additional ribose-protein binding sites in in vitro ADP-ribosylated nuclear proteins: Besides C-1″ (Hayaishi et al., Stocken et al.) C-2′ and/or C-3′ (purine-near) as well as C-2″ and/or C-3″ (pyrimidine-near), not only at the end but also within the chain of oligo-ADPR.

Zusammenfassung

Aus der Rattenleber isolierte Zellkerne werden mit NAD⊕ inkubiert, dessen beide Ribosen mit jeweils3H oder14C markiert sind. Enzymatischer (Phosphodiesterase) und/oder chemischer (Hydroxylamin) Abbau markierter ADP-ribosylierter Proteine und Identifizierung der aus Zellkernrückständen freigesetzten Bruchstücke erweist, daß AMP nach Hydroxylaminolyse sowie iso-ADP-Ribose nach Phosphodiesterase und Hydroxylamin — bei Fehlen nachweisbarer Mengen an Ribose-5-phosphat — das Vorkommen zusätzlicher Bindungsformen zwischen Protein und Ribose in in vitro ADP-ribosylierten Zellkern-Proteinen verlangen: Neben C-1″ (Hayaishi u. a., Stocken u. a.) noch C-2′ und/oder C-3′ (Purin-nahe) und weiterhin C-2″ und/oder C-3″ (Pyrimidin-nahe), und dies nicht nur am Kettenende, sondern auch in der Mitte der Kette von Oligo-ADPR.

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Author notes

  1. G. Siebert

    Present address: Division of Experimental Dentistry, University of Würzburg, Pleicherwall 2, D-8700, Würzburg, Germany

Authors and Affiliations

  1. Department of Biological Chemistry and Nutrition, University of Hohenheim, Germany

    D. Stärk &  G. Siebert

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  1. D. Stärk
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  2. G. Siebert
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L. S. Dietrich † and G. Siebert, unpublished

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Stärk, D., Siebert, G. A note on the ADP-ribose-protein linkages in rat-liver nuclei: a possible approach to assessing megavitamin therapy with niacin. Z Ernährungswiss 22, 78–84 (1983). https://doi.org/10.1007/BF02026204

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