Structural and immunological studies on the soluble formate dehydrogenase from Alcaligenes eutrophus.
During growth with formate as the sole energy source the autotrophic bacterium Alcaligenes eutrophus synthesizes a cytoplasmic formate dehydrogenase. The enzyme is a molybdo-iron-sulfur-flavo protein and the major NADH-producing system under these growth conditions, although it was estimated to constitute only 0.65% of the soluble cell protein. An electron microscopic analysis of the purified enzyme revealed that the particle is made up of four nonidentical submasses, corroborating previous structural data. The NH2-terminal amino acid sequences of the enzyme subunits exhibited significant similarities to those of only one other heteromeric formate dehydrogenase, the enzyme from the methane-utilizing bacterium Methylosinus trichosporium. Metal analyses yielded 21.5 g-atom iron, 2.18 g-atom nickel, 0.76 g-atom molybdenum, and 0.59 g-atom zinc per mol of enzyme. Initial electron paramagnetic resonance spectroscopic studies showed at least three distinct signals which appeared upon reduction of the enzyme with NADH or formate. The corresponding spin systems could be attributed to iron-sulfur centers of the enzyme. Comparative immunostaining and activity-staining experiments using cell extracts from various bacteria established immunological similarities between the soluble formate dehydrogenase of A. eutrophus and the soluble enzymes from all tested facultative autotrophs as well as from M. trichosporium.
Friedebold
J
J
Institut für Mikrobiologie, Georg-August-Universität Göttigen, Germany.
Mayer
F
F
Bill
E
E
Trautwein
A X
AX
Bowien
B
B
D000421:Alcaligenes / Q000201:enzymology* / Q000648:ultrastructure; D000595:Amino Acid Sequence; D015153:Blotting, Western; D004578:Electron Spin Resonance Spectroscopy; D005560:Formate Dehydrogenases / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D007122:Immunoelectrophoresis; D007501:Iron; D008772:Methylococcaceae / Q000201:enzymology; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D008982:Molybdenum; D013455:Sulfur
D013455:Sulfur; D008982:Molybdenum; D007501:Iron; D005560:Formate Dehydrogenases