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Lecture 23 - BIO
Lecture 23 - BIO
Pilani Campus
Lecture 23
28/02/2023
BITSPilani, Pilani Campus
Transport of metal ions in vivo
3/5/2023
BITSPilani, Pilani Campus
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
3/5/2023 BITSPilani, Pilani Campus
Iron transport
3/5/2023
BITSPilani, Pilani Campus
Overall structure of Tf
• Both iron atoms gets deeply buried within the protein structure,
which might be protecting the iron atom from siderophores
3/5/2023
BITSPilani, Pilani Campus
Iron transport
3/5/2023
BITSPilani, Pilani Campus
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
3/5/2023
BITSPilani, Pilani Campus
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
3/5/2023
BITSPilani, Pilani Campus
Iron transport
3/5/2023
BITSPilani, Pilani Campus
Stability constants of Metal-Tf complexes
3/5/2023
BITSPilani, Pilani Campus
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
3/5/2023
BITSPilani, Pilani Campus
Mechanism of Iron Binding (contd)
3/5/2023
BITSPilani, Pilani Campus
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)
3/5/2023
BITSPilani, Pilani Campus
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