Affinity chromatography on immobilised triazine dyes. Studies on the interaction with multinucleotide-dependent enzymes.
A systematic investigation into the interaction of several triazinyl dyes with two enzymes from purine metabolism, IMP dehydrogenase (IMP: NAD+ oxidoreductase, EC 1.2.1.14( and adenylosuccinate synthetase (IMP: L-aspartate ligase (GDP-forming), EC 6.3.4.4) has been conducted. Evidence from kinetic inhibition studies, enzyme inactivation with specific affinity labels and specific elution techniques from agarose-immobilised dyes indicate that triazine dyes such as Procion Blue H-B (Cibacron Blue F3G-A), Red HE-3B and Red H-3B are able to differentiate between the nucleotide-binding sites of these enzymes. This information has been exploited to design specific elution techniques for the purification of these enzymes by affinity chromatography.
D000265:Adenylosuccinate Synthase / Q000378:metabolism*; D000873:Anthracenes / Q000378:metabolism*; D001224:Aspartic Acid / Q000378:metabolism; D001665:Binding Sites; D002846:Chromatography, Affinity / Q000379:methods; D004396:Coloring Agents; D004926:Escherichia coli / Q000201:enzymology; D006160:Guanosine Triphosphate / Q000378:metabolism; D007168:IMP Dehydrogenase / Q000378:metabolism*; D007291:Inosine Monophosphate / Q000378:metabolism; D007658:Ketone Oxidoreductases / Q000378:metabolism*; D007700:Kinetics; D008025:Ligases / Q000378:metabolism*; D014227:Triazines / Q000378:metabolism*
D000873:Anthracenes; D004396:Coloring Agents; D014227:Triazines; D007291:Inosine Monophosphate; D001224:Aspartic Acid; C012842:Cibacron Blue F 3GA; C020795:procion red HE-3B; D006160:Guanosine Triphosphate; D007168:IMP Dehydrogenase; D007658:Ketone Oxidoreductases; D008025:Ligases; D000265:Adenylosuccinate Synthase