Determination of dissociation constants for binary dehydrogenase-coenzyme complexes by (Bio)affinity chromatography on an immobilized AMP-analogue

✍ Scribed by Peter Brodelius; Klaus Mosbach

• Book ID: 102982602
• Publisher: Elsevier Science
• Year: 1976
• Tongue: English
• Weight: 511 KB
• Volume: 72
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(76)90576-5

✦ Synopsis

Various alcohol and lactate dehydrogenases were adsorbed to an affinity column of immobilized W-(6-aminohexyl)-AMP and subsequently eluted with gradients of coenzymes or coenzyme fragments. A linear relation was observed between the eluting concentration of nucleotide and the reported dissociation constants for the corresponding binary enzyme-nucleotide complexes. This relation has been utilized to determine unknown dissociation constants by affinity chromatography. The method presented can also be utilized for the estimation of dissociation constants between dehydrogenases and coenzyme analogues.