Subject Chemistry

Paper No and Title 15. Bioinorganic Chemistry

Module No and Title 23. Iron storage and transport proteins

Module Tag CHE_P15_M23_e-Text

CHEMISTRY Paper No. 15: Bioinorganic Chemistry

Module No. 23: Iron storage and transport proteins
 TABLE OF CONTENTS
1. Learning Outcomes
2. Introduction
3. Iron transport by Transferrin
3.1 Role of transferrin
3.2 Structure of human serum transferrin
3.3 Mode of action
4. Iron storage
4.1 Ferritin
4.1.1 Structure of Ferritin
4.1.2 Magnetic properties of Ferritin
4.1.3 Function of Ferritin
4.1.4 How does ferritin store and release iron?
4.2 Hemosiderin
4.2.1 Magnetic properties of hemosiderin
5. Iron disorders
5.1 Iron Overload Hemachromatosis
5.1.1 Hereditary hemachromatosis
5.1.2 Transfusional Iron Overload
5.2 Iron deficiency anaemia
6. Summary

CHEMISTRY Paper No. 15: Bioinorganic Chemistry

Module No. 23: Iron storage and transport proteins
 1. Learning Outcomes
After studying this module, you shall be able to:

 Know the crucial role played by iron in various metabolic processes.

 Learn the significance of iron homeostasis i.e. why it is important to maintain a constant
balance between iron uptake, transport and storage.
 Identify the iron transport and storage proteins –“Transferrin and Ferritin respectively.”
 Study the structure, function and mode of action of Transferrin.
 Study the structure, function and magnetic properties of Ferritin and how this protein
stores and releases iron.
 Acquire information about iron related disorders such as iron overload (namely
Hereditory hemachromatosis and Transfusional Iron Overload) and iron deficiency
(anemia).

2. Introduction
Iron is a vital element that is required by all forms of life on earth as it serves as an integrated part
of many important enzymes that play a crucial role in several metabolic processes such as oxygen
transport, deoxyribonucleic acid (DNA) synthesis and electron transport. Since iron is required for
a number of diverse cellular functions, therefore it becomes extremely important to maintain a
constant balance (i.e. iron homeostasis) between iron uptake, transport, storage and utilization
processes. During the past decade, advances in our knowledge and understanding of the structure
and function of the iron-storage and transport proteins namely transferrin and ferritin have greatly
accelerated. The following sections elaborate the specific role played by each of these
metalloproteins:

3. Iron transport by Transferrin

Transferrins are group of non heme iron-binding glycoproteins that can effectively control the level
of free iron in biological fluids. Transferrin glycoproteins bind iron tightly but reversibly in the
cleft of each of two homologous lobes and hence capable of transporting it to the sites of absorption,
utilization and storage. They are widely distributed in the physiological fluids of most vertebrates
as well as invertebrates including serum, egg white, mammalian milk, tears, and leukocytes. These
transferrin families are customarily divided into following major subcatagories based on the amino
acid sequence, function and source from which each is isolated.

• Serum transferrin: Serum transferrin also known as serotransferrin, siderophilin and '13a metal-
binding globulin is generally present in the serum of vertebrates and some other biological fluids.
It plays a vital role in transport of iron within the circulatory system of the vertebrates between
different biological tissues.

• Lactotransferrin: Lactotransferrin also termed as lactoferrin is generated by mucosal epithelial

cells of mammals and therefore it is found abundantly in mammalian milk. In addition, it is also
present in specific granules of polymorphonuclear leukocytes and other secreted fluids such as
CHEMISTRY Paper No. 15: Bioinorganic Chemistry
Module No. 23: Iron storage and transport proteins
 tears, pancreatic juice etc. It plays pivotal role in body defense against infections owing to
numerous outstanding properties like antioxidant, antiinflammatory and antimicrobial activities.

• Ovotransferrin (conalbumin): Ovotransferrin or conalbumin which is commonly found in

oviduct secretions, eggs of birds and reptiles possess antimicrobial property that is vital for bird’s
innate immunity . The structural protein of ovotransferrin is similar to that of serum transferrin
since they are derived from the same gene.

• Melanotransferrin (a p97 cell surface protein): Melanotransferrin is one of the first cell surface
markers associated with melanomas. The exact biological function of melanotransferrins is
unknown, however, it has been predicted that it may play major role in cell proliferation, migration,
angiogenesis and tumorigenesis.

3.1 Role of transferrin

Transferrin is mainly produced by the liver cells which is then secreted into plasma. The main
function of transferrin is to transport iron from absorption centers to the sites of utilization, storage
and haemoglobin degradation. It also plays a key role in areas where erythropoiesis and active cell
division occur.

3.2 Structure of Human Serum Transferrin

Figure 1: Structure of human serum transferrin

Human serum transferrin is a 79.6 kDa blood plasma glycoprotein made up of polypeptide chain
containing 679 amino acids. It has a bilobal structure with two homologous lobes of almost equal
size namely amino- (N lobe) and carboxy- (C lobe) terminal lobe joined by linking polypeptide
(Figure 1). N lobe consists of amino acids 1 to 331 while C lobe is formed by amino acids 339 to

CHEMISTRY Paper No. 15: Bioinorganic Chemistry

Module No. 23: Iron storage and transport proteins
 679. Each lobe is further divided into subdomain, N1(1-92, 247-331), N2(93-246), C1(339-425,
573-679) and C2(426-572) which are interconnected by two antiparallel β strands that acts as a

hinge. Both N and C lobes are capable of binding one iron atom with the help of six ligands; Four
ligands (an aspartic acid, two tyrosines, and a histidine) are provided by the protein and two ligands
are provided by a synergistically bound carbonate anion.

3.3 Mode of action

Figure 2: Pictorial representation showing role of transferrin in the cellular iron uptake

Iron enters into cells by receptor-mediated endocytosis in a form of monoferric and diferric
transferrin (Figure 2). First of all, the iron-loaded transferrin binds to receptors present on the outer
surface of the plasma membrane. Subsequently, transferrin is internalized via invagination of
clathrin-coated pits followed by development of endocytic vesicles. After that, the clathrin is
removed with the help of uncoating enzymes and the vesicle combines with endosomes. The pH of
endosomes is reduced to about 5.5 due to the activity of V-type ATPases in their membranes which
weakens the association between iron and transferrin. This would eventually facilitates the removal

CHEMISTRY Paper No. 15: Bioinorganic Chemistry

Module No. 23: Iron storage and transport proteins
 of iron from transferrin. Transferrin along with its receptor is then transported through the endocytic
cycle back to the cell surface which becomes ready for another round of iron uptake.

4. Iron Storage
Iron that is not required in a functional capacity is typically stored in three major sites: the liver,
bone marrow and spleen. It is the iron transport protein “Transferrin” that brings iron to these sites
especially in the liver where almost 60% of body’s iron is stored while the remaining 40% is found
predominantly in the reticuloendothelial cells (RE) within the liver, spleen and bone marrow.
Further, most of the iron present in the RE cells are obtained from the phagocytosis of red blood
cells and the successive degradation of hemoglobin within the cells. The storage of iron primarily
occurs in two forms: (i) Ferritin and (ii) Hemosiderin. Initially, iron is stored as a protein-iron
complex “ferritin” however, with time, the ferritin molecules aggregate to form clusters which are
subsequently engulfed by lysosomes and consequently degraded. As a result of this process-
“hemosiderin”- an amorphous aggregate of denatured protein is formed. We shall now discuss
about the individual roles of these two important iron storage complexes:

4.1 Ferritin:

Ferritin is the principal storage form of iron in cells. It is water soluble intracellular protein found
in almost all living organisms (both prokaryotes as well as eukaryotes like mammals, plants, fungi,
bacteria) and synthesized in a variety of tissues especially within the liver, bone marrow, spleen
and intestines. A typical ferritin molecule forms a hollow sphere which contains about 2000 iron
atoms and may potentially hold upto 4500 iron atoms. The ferritin that has not been combined with
iron is commonly termed as “apoferritin.”

4.1.1 Structure of Ferritin:

The structure of ferritin is of considerable heterogeneity and comprises primarily of two important
components: a protein coat and an inorganic core suitably encapsulated within the coat. Figure 3
shows the three dimensional representation of ferritin which clearly shows that this protein
molecule is spherical in shape containing iron atoms in the form of a mineral.

CHEMISTRY Paper No. 15: Bioinorganic Chemistry

Module No. 23: Iron storage and transport proteins
 Figure 3: A three-dimensional representation of ferritin, the iron-storage protein found in our body

Structure of protein coat:

The protein coat is a highly conserved spherical assemblage made up of 24 polypeptide subunits;
each subunit consisting of about approximately 175 amino acids varying in molecular weight from
18 to 24 K Daltons. Although, the precise sequence of amino acid in the protein is not known, yet
a similar type of amino acid sequence has been detected in ferritin isolated from either animals or
plants. X-ray crystallographic study reveals that each protein subunit is folded in the form of an
ellipsoid as illustrated in Figure 4 and these ellipsoidal subunits are further packed in cubic
symmetry with fourfold, three fold and two fold axes. In the interior of the symmetrical protein
coat, there exists a hollow sphere of approximately 100 Ao diameter which is surrounded by walls
of approximately 10Ao thickness. At the intersection of the subunits, two separate channels are
found which may be filled with varying amounts of solvent and iron. These channels have a four-
fold and three fold axis respectively and allow the movement of iron in and out of the protein.
While, the four fold axis are lined with hydrophobic leucine residues, three fold axis channels are
lined with hydrophilic aspartate, glutamate and serine residues.

CHEMISTRY Paper No. 15: Bioinorganic Chemistry

Module No. 23: Iron storage and transport proteins
 Figure 4: Top view of ferritin protein coat

Structure of iron cores:

Chemically, iron in the ferritin core is stored in the form of a crystalline ferric-oxyhydroxy-
phosphate complex that has the chemical formula “FeO(PO3H2).8Fe(OH)3.” The iron cores are
unevenly arranged within the protein molecule and it is only a fraction of Fe(III) that binds directly
to this protein. As revealed by X-ray crystallographic studies, iron is present as the mineral
ferrihydrate 5Fe2O3.6H2O. Although the specific function of phosphate is not known, yet it is
thought to be involved in binding of the ferric oxide hydroxide units to each other as well as to the
protein coat.

4.1.2 Magnetic properties of Ferritin:

Every single iron atom within the ferritin molecule contains 3 unpaired electrons. The spins from
these thousands of electrons lock together under an external magnetic force, via a quantum process
known as “exchange coupling.” The core material is predominantly superparamagnetic in nature
as it acts as a solid piece of metal having single magnetic domains.

4.1. 3 Function of Ferritin:

Ferritin serves two major roles: Firstly, it provides a reserve of the element “iron” which can be
drawn upon when needed for synthesizing important iron containing proteins such as haemoglobin,
cytochromes and iron-sulphur compounds. Secondly, it protects the cell from the toxic effects of
free iron which may otherwise catalyse the activation of dioxygen leading to the formation of more
reactive oxy radicals.

4.1.4 How does Ferritin store and release Iron?

CHEMISTRY Paper No. 15: Bioinorganic Chemistry

Module No. 23: Iron storage and transport proteins
 Ferritin stores and releases iron in a controlled fashion. Since ferritin has the shape of a hollow
sphere, it stores iron the form of Fe (III). While releasing iron as and when required by the body,
Fe(III) should be changed to the Fe(II) oxidation state. Once converted to the Fe(II) form, it leaves
through the three fold and four fold channels in the spherical structure.

4.2 Hemosiderin

Hemosiderin is another iron-storage complex that is found most commonly in macrophages and
formed during breakdown of haemoglobin molecules. In contrast to ferritin, its molecular nature
remains poorly defined. It appears to be a complex of ferritin, denatured ferritin and other material.
The iron that occurs within the deposits of hemosiderin is poorly available to deliver iron when
required. However, it is abundant in situations following hemorrhage that suggests that its
formation may be related to phagocytosis of red blood cells and hemoglobin.

Figure 5: Dark hemosiderin granules in Kupffer liver cells (Prussian blue stain)

The lysosomes assimilate large amounts of hemosiderin molecules in cells that are overloaded
with iron which can be readily seen via Prussian blue staining (Figure 5).

4.2.1 Magnetic properties of Hemosiderin

Hemosiderin is also superparamagnetic in nature just like ferritin. However, in comparison to

ferritin, hemosiderin’s magnetic susceptibility effects are even more powerful owing to larger
particle size.

5. Iron disorders
It is important for us to maintain an adequate supply of iron in the body in order to synthesize
haemoglobin or other molecules that require iron for their proper functioning. Yet, because of
everyday processes like urination, sweating, defecation etc, our bodies keep continually losing iron
in small quantities. For compensating these losses, one must consume approximately 18 mg of iron
every day. Also, there are instances wherein too much of iron is deposited in the body tissues that
leads to a situation called “iron overload.” Following sections throw light on iron related disorders:

CHEMISTRY Paper No. 15: Bioinorganic Chemistry

Module No. 23: Iron storage and transport proteins
 5.1 Iron Overload (Hemochromatosis):

An iron overload disorder which makes the body absorb excessive amounts of iron from the food
and drink we consume is often termed as “Hemochromatosis.” Total body iron overload
(Hemochromatosis) occurs most often either due to “hereditary hemochromatosis” or “repeated
transfusions in patients suffering from severe anaemia. Hereditary hemochromatosis- “an
autosomal recessive disorder” is the more common of the two by far.

5.1.1 Hereditary hemachromatosis:

Hereditary haemochromatosis (HH) is a genetic disorder that reflects a fractional increase in dietary
iron absorption. Under normal situations, humans extract iron from food through the intestines and
once iron is present in sufficient amount, the body reduces its absorption to avoid excessive
accumulations. However, in case of people suffering from hereditary hemochromatosis, the body
absorbs too much of iron and in due course (i.e. over several years) this amount may increase upto
5 to 20 times. This excess iron the body gets stored in the tissues of our major organs such as liver,
the heart and pancreas which may eventually destroy these organs, causing severe chronic diseases
such as cirrhosis, heart disease and diabetes. The gene chiefly responsible for haemochromatosis-
“HFE” (identified on chromosome 6) encodes a protein that is homologous to the Class I HLA
antigens. An alteration in this HFE protein basically involves the mutation of a cysteine to a leucine
at position 282 (C282Y). People who inherit one copy of the mutant HFE gene are carriers who
rarely develop iron overload and as such do not show any symptoms since one correct copy
regulates the iron absorption process (Figure 6). However, people with two inherited copies of the
mutant protein may develop this disease and consequently face a myriad of problems because of
this. Almost 90% of people with hereditary haemochromatosis have the C282Y mutation in HFE.
The most common symptoms of this inherited iron overload disorder include Fatigue, weakness
and lethargy, Joint pains leading to osteoarthritis etc.

CHEMISTRY Paper No. 15: Bioinorganic Chemistry

Module No. 23: Iron storage and transport proteins
 Figure 6: Diagram representing the genetic inheritance of this disorder

5.1.2 Transfusional Iron Overload

This iron overload disorder occurs in case of transfusion dependant patients like those suffering
from thalassemia major, sickle cell disease, myelodysplastic syndrome, aplastic anemia, hemolytic
anemia, refractory sideroblastic anemias and diamond-blackfan anemia. Because of the excess iron
received during each transfusion and because there is no physiological means of excreting it, this
element gradually accumulates in the organs of such patients particularly in the liver. The
assimilation of iron with repeated transfusion shows that the heme iron atoms from the transfused
red cells are retained after they have become senescent. The senescent red cells are ultimately
destroyed by the reticuloendothelial cells (RE cells). Although the pattern of organ injury is similar
in both transfusional iron overload as well as hereditary hemochromatosis, however the
distinguishing feature between the two is the occurrence of large iron deposits in the RE cells in
case of the former.

5.2 Iron deficiency anaemia

Iron is very important for the production of a globular protein called hemoglobin which is
responsible for carrying oxygen from the lungs to various body tissues for respiration to provide
energy to power the functions of the organism. Hence, the deficiency of iron in blood stream leads
to decrease in hemoglobin level because of which the organs will not get enough amount oxygen
and such condition is termed as anaemia (Figure 7). It occurs due to insufficient dietary intake of
iron, or iron loss from bleeding either intestinal, uterine or urinary tract. Iron-deficiency anemia
can lead to fatigue, shortness of breath, chest pain and heart problems.

CHEMISTRY Paper No. 15: Bioinorganic Chemistry

Module No. 23: Iron storage and transport proteins
 Figure 7: Diagram representing iron deficiency anemia

5. Summary
 Iron is an essential element for almost all living organisms since it plays a crucial
role in a number of diverse cellular functions. Therefore, it becomes extremely important
to maintain a constant balance between iron uptake, transport, storage and utilization
processes.
 Transferrins which are widely distributed in the biological fluids of most vertebrates as
well as invertebrates are group of non heme iron-binding glycoproteins that are capable of
transporting iron from absorption centers to the sites of utilization, storage and
haemoglobin degradation.
 The storage of iron primarily occurs in two forms: (i) Ferritin and (ii) Hemosiderin.
Initially, iron is stored as a protein-iron complex “ferritin” however, with time, the ferritin
molecules aggregate to form clusters which are subsequently engulfed by lysosomes and
consequently degraded. As a result of this process- “hemosiderin”- an amorphous
aggregate of denatured protein is formed.
 An iron overload disorder which makes the body absorb excessive amounts of iron from
the food and drink we consume is often termed as Hemochromatosis. Total body iron
overload occurs most often either due to “hereditary hemochromatosis” or “repeated
transfusions in patients suffering from severe anaemia.
 The deficiency of iron in blood stream leads to decrease in hemoglobin level because of
which the organs will not get enough amount oxygen and such condition is termed as
anemia.

CHEMISTRY Paper No. 15: Bioinorganic Chemistry

Module No. 23: Iron storage and transport proteins