Iron metabolism of Escherichia coli studied by Mössbauer spectroscopy and biochemical methods.
To date it has barely been recognized that the nature of about 75% of the Escherichia coli iron pool is unknown. Here we report the isolation of two iron species representing major components of iron metabolism in various growth states of E. coli. In vivo Mössbauer spectroscopy was applied to obtain information on the intracellular distribution pattern of iron in E. coli K12 W3110. Only two types of iron could be detected in the cell spectra: hexacoordinated Fe2+ and Fe3+ high-spin complexes. Other iron-requiring compounds are at least one order of magnitude less abundant in E. coli. The Mössbauer parameters of these complexes fit neither cytochromes nor iron-sulfur proteins nor ferric holo-bacterioferritin. They are sensitive to metabolic changes and inhibitors. The ratio of Fe/subunit, Fe2+/Fe3+ interconversion, chromatographic and electrophoretic data exclude bacterioferritin as the main iron metabolite in E. coli. Bacterioferritin can be observed only at very high ferric ion concentrations in the medium. The 55Fe fluorograms of both cytoplasmic and membrane fractions exhibit two exclusive bands with apparent molecular masses of 17 and 15 kDa, respectively. The two bands comprised 70% of the applied radioactivity. In gel filtration the main iron peak elutes at 155 kDa yielding two bands with apparent molecular masses of 17 and 15 kDa on SDS/PAGE. We therefore conclude that the iron species form a protein with an apparent molecular mass of 155 kDa containing 17-kDa and 15-kDa subunits. The iron content of the protein is 44 micrograms Fe/mg protein which corresponds to approximately 13 iron ions/subunit. No iron protein exhibiting the observed features has been described so far. Additional Mössbauer experiments suggest that these novel iron proteins are not restricted to E. coli but that similar components are detectable in several bacterial and fungal systems, thus pointing to a general occurrence.
Matzanke
B F
BF
Universität Tübingen, Institut für Biologie II, Mikrobiologie I, FRG.
Müller
G I
GI
Bill
E
E
Trautwein
A X
AX
D001426:Bacterial Proteins / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002850:Chromatography, Gel; D003573:Cytochrome b Group / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D005290:Ferric Compounds / Q000378:metabolism; D005291:Ferrichrome / Q000031:analogs & derivatives / Q000378:metabolism; D005293:Ferritins / Q000378:metabolism; D005296:Ferrous Compounds / Q000378:metabolism; D005470:Fluorometry; D007501:Iron / Q000378:metabolism*; D008970:Molecular Weight; D009997:Osmotic Pressure; D015204:Spectroscopy, Mossbauer
D001426:Bacterial Proteins; D003573:Cytochrome b Group; D005290:Ferric Compounds; D005296:Ferrous Compounds; D005291:Ferrichrome; C021678:ferricrocin; D005293:Ferritins; C018032:bacterioferritin; D007501:Iron