Enzyme complex which couples glycerol-3-phosphate dehydrogenation to fumarate reduction in Escherichia coli.
Anaerobic growth of Escherichia coli on glycerol as carbon source and fumarate as hydrogen acceptor results in the induction of both anaerobic sn-glycerol-3-phosphate (G3P) dehydrogenase and fumarate reductase. In previous studies, the G3P dehydrogenase was measured by the G3P-dependent reduction of a tetrazolium dye mediated by phenazine methosulfate in the presence of flavine mononucleotide and flavine adenine dinucleotide, and fumarate reductase was measured by the fumarate-dependent oxidation of reduced flavine mononucleotide. Results from the present study indicate that these two enzymes actually constitute a functional complex which is sedimentable at 200,000 x g in 2 h and which can catalyze the dehydrogenation of G3P at the expense of fumarate without any added cofactor. This coupling activity is not constituted by simply mixing an extract containing anaerobic G3P dehydrogenase with another extract containing fumarate reductase. Additional evidence for an organized complex is provided by the high degree of sensitivity of the coupled reaction to low concentrations of the detergents Triton X-100 and Emasol 1130 in comparison with the individual activities of the two component enzymes measured with the aid of artificial cofactors.
D000693:Anaerobiosis; D002247:Carbon Isotopes; D002474:Cell-Free System; D004926:Escherichia coli / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D005486:Flavin Mononucleotide / Q000378:metabolism; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D005650:Fumarates / Q000378:metabolism*; D005993:Glycerolphosphate Dehydrogenase / Q000378:metabolism*; D005994:Glycerophosphates / Q000378:metabolism*; D007930:Leucine / Q000378:metabolism; D009097:Multienzyme Complexes / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism*; D013053:Spectrophotometry; D013501:Surface-Active Agents / Q000494:pharmacology
D002247:Carbon Isotopes; D005650:Fumarates; D005994:Glycerophosphates; D009097:Multienzyme Complexes; D013501:Surface-Active Agents; D005182:Flavin-Adenine Dinucleotide; D005486:Flavin Mononucleotide; D010088:Oxidoreductases; D005993:Glycerolphosphate Dehydrogenase; D007930:Leucine