Inexpensive purification of P450 reductase and other proteins using 2',5'-adenosine diphosphate agarose affinity columns

Protein Expr Purif. 2001 Jun;22(1):82-3. doi: 10.1006/prep.2001.1385.

Abstract

Two reductases, P450 oxidoreductase and P450Bm-3 reductase, were purified on a 2',5'-adenosine diphosphate solid support. Although the efficiency of these columns is well established, the cost of the resin and the eluting material 2'-adenosine can be prohibitive. Herein we show that the less costly 2',3'-adenosine monophosphate is an excellent eluting material.

MeSH terms

  • Adenosine Diphosphate / metabolism
  • Adenosine Monophosphate / metabolism
  • Bacterial Proteins*
  • Chromatography, Affinity*
  • Chromatography, Agarose*
  • Cytochrome P-450 Enzyme System / isolation & purification*
  • Cytochrome P-450 Enzyme System / metabolism
  • Mixed Function Oxygenases / isolation & purification*
  • Mixed Function Oxygenases / metabolism
  • NADPH-Ferrihemoprotein Reductase / isolation & purification*
  • NADPH-Ferrihemoprotein Reductase / metabolism
  • Protein Binding

Substances

  • Bacterial Proteins
  • adenosine 2',5'-diphosphate
  • Adenosine Monophosphate
  • Adenosine Diphosphate
  • Cytochrome P-450 Enzyme System
  • Mixed Function Oxygenases
  • NADPH-Ferrihemoprotein Reductase
  • flavocytochrome P450 BM3 monoxygenases