BITS Pilani

Pilani Campus

CHEM F343: Inorganic Chemistry III

Lecture 24
02/03/2023
BITSPilani, Pilani Campus
UV-visible specral 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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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
Metal Storage

• In mammals, iron is stored in liver, spleen and bone morrow

• An average person contains 3-4 gm of iron
• major part of this is hold as storage and only 35 mg is used biological
process like oxygen transport, redox and enzymatic reactions
• Ferritins are a class of iron storage proteins found widely distributed
among the animal, plant and microbial kingdoms
• These proteins consist of 24 subunits assembled into hollow
spherical structure within which iron is stored as a hydrous ferric
oxide mineral core
• The core diameter lies between 60-80 Å
• The storage capacity of ferritin is approximately 4500 Fe3+ ions per
molecule
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Ferritin
• Funnel shaped channel that
possess a threefold axis of
symmetry
• These are lined with
hydrophilic residues (Asp,
Glu, Ser) (hydrophilic)
• Another series of channels
possesses a fourfold axis of
symmetry and are lined with
12 hydrophobic leucine
residues (hydrophobic)
• These hydrophobic channels
may provide access for
organic reductants (NADH,
FADH) or chelating agent Representative subunit showing the bundle
of helices that form the major portion of
backbone
Representation of Ferritin
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Ferritin

• Ferritin possesses eight funnel-shaped hydrophilic channels

• Channels are lined with hydrophilic residues like Asp, Glu and Ser
• Ferritin core is composed of Ferric oxyhydroxy phosphate complex
[FeO(PO3H2).8Fe(OH)3]
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Ferritin (contd)

• Why does Ferritin takes up iron as divalent metal?

 Fe(III) is insoluble at intracellular pH
 Fe(II) is kinetically labile relative to Fe(III)
(Fe(III) (kex (Fe(II) ~ 107 s-1; kex(Fe(III) ~ 103s-1) which is essential for
rapid complexation
 Inertness of Fe(III) facilitates for long term storage or transport

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 Metalloproteins and Metalloenzymes:
Oxygen Carriers and Hydrolases

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Oxygen Carrier
• Transition metal ions have been extensively utilized by nature in the
design of metalloprotein and enzymes
• The metals are involved in wide variety of functions (catalysis;
storing; transferring, regulating the activities of biomolecules)
• Selection and specificity of transition metals does not simply
depends on ionic radius like alkali and alkaline earth metal ions
• Combination of ligand preference and geometric constraints is used
to differentiate both metal ions and oxidation state (see the Table)
• Transition metals are most often associated with oxygen binding and
oxido-reductase protein/enzymes
• Transition metal ions can also act as Lewis acid
• Ex.: Ni(II) in urease, Fe(III) in acid phosphatase, [Fe4S4]2+ in
hydrolyases
• Role of Lewis acid is commonly fulfilled by Mg and Zn
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Oxygen carrier
• Changing of reducing environment of earth into oxidizing one
resulted in the evolution of O2 binding proteins
• Proliferation of cyanobacteria ~2-2.5 billion y ago resulted in the
change through photosynthetic process capable of decomposing
water with the release of oxygen
• O2 should not easily diffuse in muscle as it is toxic to biological
systems, so organisms have developed a way for it’s transportation
• Three known O2-binding system, heme ligand, binuclear iron and
copper complexes developed
• e-transport protein are found in lower forms of organisms; thus it is
predicted these O2-transport and storage proteins evolved from
these e-transport protein (similar prosthetic group)

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