BITS Pilani

Pilani Campus

CHEM F343: Inorganic Chemistry III

Lecture 23
28/02/2023
BITSPilani, Pilani Campus
Transport of metal ions in vivo

• In multicellular organism, transportation of minerals and

nutrients from a point of ingestion to a location where
metal ion gets used or stored.
• Minerals and nutrients derived from food and fluids gets
absorbed into the blood stream.
• The alkali (and to some extent alkaline earth) metals are
soluble under physiological pH and do not interacts strongly
or irreversibly with ligating groups on cell walls or
membrane
• These ions can be transported as free ions in the blood
stream and do not require special carrier proteins or ligands
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Transport of trace transition metal ions
• At physiological O2 concentration, Fe2+ is readily oxidized to Fe3+,
which is highly insoluble in aqueous solution at neutral pH
Fe3+ (aq) + 3OH- (aq) Fe(OH)3 (s) Ksp ~ 10-39 M4
• To overcome such serious solubility problem, trace transition metals
require special carrier proteins to bind and transport the ions in vivo

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Iron transport
• Iron transport is most thoroughly investigated
• Firstly, it must be transported from the food in the gut to the places
where it is required.
• Mostly, iron is required in the bone marrow, where red blood cells
are formed.
• Iron cannot be transported around the body's circulation system as
free iron, since it would be susceptible to chelation by siderophores,
or may precipitate as iron(III) oxide, or may form iron(II)
• After entering the mammalian bloodstream by passage through
intestinal membrane, free iron is captured by protein called
transferrin (Tf)
• Transferrins are a class of nonheme iron binding protein with a
relative molecular mass of about 80 000
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Iron transport

• They include serum transferrin, ovotransferrin, lactoferrin and

melanotransferrin
• lactoferrin is found in milk, while Ovotransferrin in egg white
• Tf isolated from blood plasma is occasionally referred as
serotransferrin while Ovotransferrin as Conalbumin
• Tf are glycoproteins, that bind two Fe(III) ions in similar but
structurally distinct sites termed as N-terminal and C-terminal
domains

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Overall structure of Tf

• Both iron atoms gets deeply buried within the protein structure,
which might be protecting the iron atom from siderophores

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Iron transport

• Carbonate ion is required for strong

Fe3+ coordination
• Neither Fe3+ nor CO32- binds Tf strongly
on its own, but together binding favours
• Such cooperative behavior is termed
as synergism
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Features of Tf during iron binding
• The iron binding site is rich in hard ligands, which are suitable for
binding iron(III)
• When the iron atom enters the active site, it is coordinated by one
η1-aspartyl, one histidyl and two tyrosinate side-chains; a non-protein
ligand carbonate (CO32−)
• Carbonate group is held in place within the protein via hydrogen
bonding interactions
• The carbonate coordinates to the iron in an η2 fashion; in other
words, it is a chelating ligand.
• It is not clear why this unusual synergistic binding of iron and
carbonate occurs in transferrin, but it may have something to do with
the way iron is released from transferrin.

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UV-visble specrrral studies of Fe-Tf

(a) Tf at pH 6
(b) Fe-Tf at pH 6
(c) Tf at pH 11
(d) Fe-Tf at pH 11

•Tf exhibits two absorption bands in

UV-VIS absorption due to Tyr (245
and 295nm)
•Fe binding enhances the absorption
• Red coloration of Ferritin
Fe(III) + Tf + CO32- → [Fe(III)Tf(CO32-)] + 3H+

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UV-visible studies of Fe-Tf
To understand the metal – binding by optical spectroscopy

• Titration of Conalbumin
(Tf) with Fe(III)
• Change in absorbance
monitored at 245 nm
• Reveals the binding of
two iron/Tf

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Iron transport

• Since all the coordinating atoms in transferrin is hard in nature, Tf

forms stable complex with iron(III).
• The stability constant of the Fe(III)–transferrin complex is in the
order of 1020 M−1
• Strong stability constant protects the iron(III) against the low
concentration of any siderophores present
• Transferrins show mild antibacterial properties to prevent extensive
iron chelation by siderophores
• Transferrin also forms relatively stable complexes with other hard
metals

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Stability constants of Metal-Tf complexes

Metal Log10(stability constant)

Cadmium(II) 5.95
Zinc(II) 7.80
Aluminium(III) 13.50
Iron(III) 22.80

• These data suggests that other metals can also be transported by

transferrin into cells, where they are potentially harmful.
• Aluminium, is of major concern because it is used widely in cooking
utensils
• Accumulation of Aluminium leads to Alzheimer's disease

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Mechanism of Iron Binding

• These macroscopic
binding constants provide
no indication of the
distribution of iron
• Individual constants can
be obtained by gel
electrophoresis
experiment
• N-terminal Tf releases
iron at lower pH (~5.7),
while C-terminal retains it

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Mechanism of Iron Binding (contd)

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Iron translocation by Tf

• Endocytosis: Process by
which cell breaks off to
form an internal vesicle
• Membrane bound H+-
ATPase pumps the
protons into the vesicle
to drop the pH by 5.5 to
release iron as Fe(II)

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Determination of Intracellular vs extracellular pH
Determination of pH at extracellular:
At 0°C the Tf(Fe(III)2 complex binds to
the surface receptor, endocytosis does
not occur; so the dye emission is
determined by the extracellular pH

At 37°C, the complex is taken into the cell,

and the lower intracellular pH is reflected
in the emission profile