FAD requirement for the reduction of coenzyme F420 by hydrogenase from Methanobacterium formicicum.
Hydrophobic interaction chromatography of coenzyme F420-reducing hydrogenase purified from Methanobacterium formicicum depleted protein-bound FAD and eliminated the ability to reduce coenzyme F420. Preincubation of the FAD-depleted hydrogenase with FAD restored 85% of the coenzyme F420-reducing activity. FMN did not replace FAD. A Kd of 12 microM was estimated for FAD. Analysis of the reactivated hydrogenase following molecular sieve column chromatography showed that FAD was bound to protein. The results indicate that protein-bound FAD is reversibly removed from the coenzyme F420-reducing hydrogenase and that this flavin is required for the reduction of coenzyme F420.
D002845:Chromatography / Q000379:methods; D019605:Euryarchaeota / Q000201:enzymology*; D005182:Flavin-Adenine Dinucleotide / Q000502:physiology*; D006864:Hydrogenase; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D011485:Protein Binding; D012256:Riboflavin / Q000031:analogs & derivatives* / Q000378:metabolism
D005182:Flavin-Adenine Dinucleotide; C015084:coenzyme F420; D010088:Oxidoreductases; D006864:Hydrogenase; D012256:Riboflavin