The purification of nicotinamide nucleotide-dependent dehydrogenases on immobilized cofactors

Biochem J. 1973 Jul;133(3):507-13. doi: 10.1042/bj1330507.

Abstract

1. The general applicability of affinity chromatography to the purification of nicotinamide nucleotide-dependent dehydrogenases on immobilized cofactors is illustrated with several examples taken from crude systems. 2. Methods for overcoming the inevitable loss of selectivity experienced with these polymers are suggested. Effective use of the appropriate nucleotide, the second substrate and other interacting ligands can be made to selectively alter the chromatographic behaviour of the desired enzyme.

MeSH terms

  • Adenosine Diphosphate
  • Adenosine Monophosphate
  • Aminocaproates
  • Chromatography*
  • Chromatography, Affinity
  • Glucosephosphate Dehydrogenase
  • NAD
  • NADP
  • Niacinamide*
  • Oxidoreductases / isolation & purification*
  • Polysaccharides
  • Pseudomonas / enzymology
  • Rhodobacter sphaeroides / enzymology
  • Ribonucleotides*
  • Saccharomyces cerevisiae / enzymology
  • Threonine

Substances

  • Aminocaproates
  • Polysaccharides
  • Ribonucleotides
  • NAD
  • Niacinamide
  • Threonine
  • Adenosine Monophosphate
  • NADP
  • Adenosine Diphosphate
  • Oxidoreductases
  • Glucosephosphate Dehydrogenase