Abstract
Recent studies have indicated that Escherichia coli possesses at least
two iron-storage proteins, the haem-containing bacterioferritin and
ferritin. The ferritin protein has been amplified 600-fold to 11-14%
of total cell protein in a bfr mutant and purified to homogeneity
with an overall yield of 13%. The cellular ferritin content remained
relatively constant throughout the growth cycle and amplification
was accompanied by a 2.5-fold increase in cellular iron content.
The isolated ferritin contained 5-20 non-haem iron atoms/holomer
and resembled the eukaryotic ferritins rather than the prokaryotic
bacterioferritins in containing no haem. The 24 subunits of this
ferritin (M(r) 19400) assemble into a spherical protein shell (12
+/- 1 nn diameter, M(r) 465 000) which sequesters at least 2000 iron
atoms in vitro to form an electron-dense iron core of 7.9 +/- 1 nm
diameter. Electron-microscopic and Mossbauer spectroscopic studies
with iron-loaded ferritin showed that the core can be either crystalline
(ferrihydrite) or amorphous, depending on the absence or presence
of phosphate, respectively. Mossbauer spectroscopy with intact E.
coli revealed a novel-high spin Fe(II) component which is enhanced
in bacteria amplified for ferritin but not in the parental strain.
Western blotting showed that ferritin and bacterioferritin are immunologically
distinct proteins. E. coli is thus an organism containing both a
ferritin and a bacterioferritin and the relative roles of the two
iron-storage proteins are discussed in this study.
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