Identification of molybdoproteins in Clostridium pasteurianum.
Cells of Clostridium pasteurianum whose N source is switched from NH3 to N2 accumulate large amounts of molybdenum beginning 1.5 h before the detection of nitrogenase activity. Anaerobic multiphasic gel electrophoresis and anion-exchange chromatography were used to identify the molybdoproteins and molybdenum-containing components present in N2-fixing cells. In addition to molybdate, six distinct 99Mo-labeled species were detected, i.e., a membrane fragment, the MoFe protein of nitrogenase, formate dehydrogenase, a Mo "binding-storage" protein, a 30-kilodalton molybdoprotein, and a low-molecular-weight molybdenum species. Of these, the MoFe protein, formate dehydrogenase, and the Mo binding-storage protein were present in more than one zone because of complex formation with other proteins, partial denaturation, and variation in the amount of Mo bound to the protein, respectively. In addition to the six proteins, a soluble "free" Mo cofactor in the cytosol was detected by showing that it reconstituted nitrate reductase activity in crude extracts of the Neurospora crassa mutant nit-1.
D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000378:metabolism; D003013:Clostridium / Q000378:metabolism*; D003067:Coenzymes; D005560:Formate Dehydrogenases / Q000378:metabolism; D008667:Metalloproteins / Q000378:metabolism*; D008970:Molecular Weight; D008982:Molybdenum / Q000378:metabolism*; D000090006:Molybdenum Cofactors; D009591:Nitrogenase / Q000378:metabolism; D011621:Pteridines / Q000378:metabolism; D013431:Sulfates / Q000378:metabolism
D001426:Bacterial Proteins; D003067:Coenzymes; D005560:Formate Dehydrogenases; D008667:Metalloproteins; D008982:Molybdenum; D000090006:Molybdenum Cofactors; D009591:Nitrogenase; D011621:Pteridines; D013431:Sulfates; C022261:molybdopterin