A two-step synthesis of new water-soluble polymers of NAD+ and ADP. The biological properties of these polymers.
Alkylation at the N-1 position of the adenine moiety of NAD+, ADP or ATP with 2,3-epoxypropyl acrylate, followed by polymerization with or without acrylamide at pH 8, gave water-soluble polymers of NAD+ and ADP where the alkyl chain was located at the exocyclic adenine C-6 amino group. Cofactor incorporations were good to high: 145-447 mumol NAD+/g polymer and 667 mumol ADP/g polymer. About 30% of the bound NAD+ could be reduced with rabbit muscle lactae dehydrogenase, yeast alcohol dehydrogenase and Bacillus subtilis alanine dehydrogenase; 84% of the bound ADP was phosphorylated with rabbit muscle creatine kinase. High cofactor activities were obtained with polymerized NAD+ with alcohol dehydrogenase as enzyme: the initial rate of NAD+ polymer reduction was 35-81% that of free NAD+. These values remained substantially high with agarose-immobilized alcohol dehydrogenase (15-36%) and should eventually allow their use in continuous enzymatic reactors. Enzymatic phosphorylation of ADP polymer by creatine kinase gave an ATP polymer with high biological activity: 480 mumol ATP/g polymer were transformed with yeast hexokinase.
D000180:Acrylic Resins; D000244:Adenosine Diphosphate / Q000031:analogs & derivatives*; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives*; D000429:Alcohol Oxidoreductases / Q000378:metabolism; D055598:Chemical Phenomena; D002621:Chemistry; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009243:NAD / Q000031:analogs & derivatives*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D010770:Phosphotransferases / Q000378:metabolism; D011108:Polymers; D011485:Protein Binding; D013329:Structure-Activity Relationship
D000180:Acrylic Resins; D011108:Polymers; D009243:NAD; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000429:Alcohol Oxidoreductases; D007770:L-Lactate Dehydrogenase; D009247:NADH, NADPH Oxidoreductases; D010770:Phosphotransferases