Iron uptake byBifidobacterium thermophilum protoplasts |
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Authors: | Eva Kot Robin Miller-Catchpole Anatoly Bezkorovainy |
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Institution: | (1) Department of Biochemistry, Rush-Presbyterian-St. Luke's Medical Center, 60612 Chicago, IL |
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Abstract: | Protoplasts ofBifidobacterium thermophilum were prepared by a combination of lysozyme and protease digestion, and ferrous iron uptake studies were carried out. Little,
if any, iron was internalized by the protoplasts, although large amounts of iron were bound to the protoplast surface. This
binding was much greater than that of intact cells, which prefer to internalize iron by an energy-dependent process. It was
also found that the binding of iron by protoplasts of cells grown in an iron-deficient medium was much more extensive than
that of cells grown in an iron-sufficient medium. Soluble and particulate fractions of protoplasts were prepared by grinding
them in a glass homogenizer, and the particulate fraction was also subjected to iron binding studies. The amount of iron bound
was the same as that in intact protoplasts, indicating that the particulate fraction membrane fragments bound iron on their
outer surface only. Nevertheless, when iron-preloaded cells were protoplasted and their surface cleared of iron, their particulate
fraction contained considerable amounts of iron, indicating that the inner surface of the membranes is capable of binding
iron as long as the cell is intact. The amount of iron so bound was dose-dependent on the amount of iron entering the cell.
The failure of the outer and inner surface iron pools to mix was confirmed by the fact that when iron-preloaded protoplasts
were incubated with additional iron, only the latter (surface-bound) was elutable with nonradioactive 2 mM FeSO4. It is concluded that increasing bifidobacterial iron load increases the amount of iron bound to the inner surface of the
membrane; the procedure, which is effective in forming bifidobacterial protoplasts, destroys their iron transport mechanism
while uncovering surface iron-binding sites; and that such iron-binding sites may be of significance in the cellular iron
metabolism processes. |
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Keywords: | Bifidobacterium thermophilum iron ferrous protoplasts |
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