Functions of Iron- Overview

e
-
donor and acceptor



Enzyme-Fe
2+
+ substrate Enz-Fe
3+
+ product ( plus 1 e
-
)

Enzyme- Fe
3+
+ substrate Enz-Fe
2+
+ product (minus 1 e
-
)



Three Fe enzyme flavors- heme, Fe-S, non-heme
Uses of Iron I - Heme Proteins
Oxygen Carriers
Hemoglobin, myoglobin

Electron Transfer (ETC/ox phos)
Cytochromes a, b, c
(convert NADH, FADH2 into ATP energy)

Metabolism of O
2
& Peroxides (R-OOH)
Cytochrome P450 (drug metabolism)
Catalase (degrades H
2
O
2
)
NADPH oxidase (phagocytes)


Uses of Iron II
Iron-Sulfur Cluster Proteins
Redox enzymes
Succinate dehydrogenase (TCA)
Xanthine oxidase (purine degradation)
Non-redox Enzymes
Aconitase (TCA)


Monooxygenases
Add OH to aromatic amino acids (neurotransmitter synth)
tyrosine dopamine
tryptophan serotonin

Dioxygenases
Synthesis of OH-lysine, OH-proline in collagen
Used to crosslink collagen fibrils in connective tissue

Enzymes containing Fe-O-Fe
Ribonucleotide Reductase (makes deoxyribose)
Uses of Iron III
Single Fe-Containing Enzymes
Iron toxicity
Insoluble in body
Fe
+3
(ferric) 10
-17
M; Fe
+2
(ferrous) 10
-9
M

Strong oxidizing agent (Fenton reaction)
Fe
+2
+ H
2
O
2
Fe
+3
+ OH + OH-
Fe
+3
+ O
2
-
Fe
+2
+ O
2
(& repeat)

Must be kept protein bound to keep it soluble &
reduce its toxicity

Toxicity risk- hemochromatosis
Fe in serum
Regulated tightly by Fe status

Transferrin is major carrier in the blood

Cell must have Transferrin Receptor to take up & use Fe

Normal status: Tf is 30% saturated with Fe

Iron also found as serum ferritin

Transferrin
pH 5.5
A) Uptake
(TfR cycle)
C) Storage B) Metabolic
Utilization
Extracellular
Space
Cytoplasm
D) Export
Heme
Iron Containing
Proteins
Ferritin
Protoporphyrin IX
5-Aminolevulinate
Succinyl-CoA + Glycine
Mitochondrion
The transferrin cycle
Dmt1
pH 5.5
A) Uptake
(TfR cycle)
C) Storage B) Metabolic
Utilization
Extracellular
Space
Cytoplasm
D) Export
Heme
Iron Containing
Proteins
Ferritin
Protoporphyrin IX
5-Aminolevulinate
Succinyl-CoA + Glycine
Mitochondrion
The transferrin cycle
Dmt1
Iron Stores
Major tissue sites are liver & bone marrow/spleen

Storage varies w/gender, age, diet, disease

Storage form: ferritin
Cytosolic multimeric protein

Reversible storage form
Ferritin
x 24
Regulation of cellular iron metabolism

Post-transcriptional control
mRNA stability
translation

Regulated by the Iron Regulatory Proteins (IRPs)
Iron metabolism

Functions and toxicity of Fe

Fe in serum- transferrin

Cellular Fe metabolism

Regulation of cellular iron

Intestinal iron absorption

Macrophage iron metabolism

The Fe cycle

Systemic control of iron metabolism- hepcidin

Hemochromatosis
Iron in the diet
Heme and nonheme iron
meats- primarily heme iron
vegetables- primarily non-heme iron

Bound to proteins

Released by proteases (e.g. pepsin) and HCl

Heme is taken up intact

Non-heme iron is transported as Fe(II)
apical basolateral
Intestinal iron uptake
Fe
2+



Fe
3+

e-
DcytB
Dmt1
Fe
2+




Hcp1
Heme Heme
Ferritin
Ferroportin
Hephaestin
Fe
2+
Fe
3+ Transferrin
Zn, Cu, and Fe Papers
1. Components of zinc transport.
Liuzzi et al. 2005, Eide 2006

2. Levels and trafficking of intracellular metal ions.
Outten and OHalloran, 2001

3. Copper transporters and their function.
Puig and Thiele 2002, Nose et al. 2006

4. Regulation of mineral homeostasis-regulated protein trafficking.
Petris et al. 2002

5. Use of model systems (e.g. yeast, nematodes, zebrafish) to identify
mammalian iron transporters.
Hentze et al. 2004, Donovan et al. 2000

6. Hormonal regulation of iron status- Hepcidin.
Nemeth et al. 2004
Fig. 1. The phenotype of
weissherbst (weh)

a. Reduced hemoglobin

b. Hypochromic anemia

c. Not due to globin
protein defects
Fig. 2. The protein product
of weissherbst (weh)-
ferroportin1
Fig.3. Ferroportin1 is an
iron exporter protein
Fig. 4. Expression of the
mammalian ferroportin
protein
apical basolateral
Intestinal iron uptake
Fe
2+



Fe
3+

e-
DcytB
Dmt1
Fe
2+




Hcp1
Heme Heme
Ferritin
Ferroportin
Hephaestin
Fe
2+
Fe
3+ Transferrin
Macrophage iron metabolism
Ferroportin
Cerulo-
plasmin
Fe
2+
Fe
3+ Tf
Fe
Ferritin
Fe
Heme
Iron metabolism

Functions and toxicity of Fe

Fe in serum- transferrin

Cellular Fe metabolism

Regulation of cellular iron

Intestinal iron absorption

Macrophage iron metabolism

The Fe cycle

Systemic control of iron metabolism- hepcidin

Hemochromatosis
Wheres the Iron in Body?
(~3 gm total)
Hemoglobin 2000 mg
Storage Fe 1000 mg
Tissue Fe
Myoglobin Fe 130 mg
Enzyme Fe 8 mg
Transport Fe (blood) 3 mg
Values are for a 70 kg adult male; female values lower
Zn, Cu, and Fe Papers
1. Components of zinc transport.
Liuzzi et al. 2005, Eide 2006

2. Levels and trafficking of intracellular metal ions.
Outten and OHalloran, 2001

3. Copper transporters and their function.
Puig and Thiele 2002, Nose et al. 2006

4. Regulation of mineral homeostasis-regulated protein trafficking.
Petris et al. 2002

5. Use of model systems (e.g. yeast, nematodes, zebrafish) to identify
mammalian iron transporters.
Hentze et al. 2004, Donovan et al. 2000

6. Hormonal regulation of iron status- Hepcidin.
Nemeth et al. 2004
Systemic control of iron status

Systemic regulators:
1. Stores
2. Erythroid
3. Hypoxia
4. Inflammatory

Sites of action
control intestinal uptake
control release of iron from macrophages

Hepicidin

Previous hepcidin studies

1. Hepcidin is a 25 amino acid serum peptide
2. Hep KO have increased Fe accumulation
3. Hep overexpressors have decreased Fe
4. Hepcidin is expressed in liver
5. Hepcidin expression is induced by:
iron
infection & inflammation (IL-6)
hypoxia
erythropoiesis

Park et al. JBC 276: 7806
C B A
Peptide sequencing EST
Hereditary hemochromatosis
Disease of Fe overload
Majority caused by mutation in HFE gene
Most common genetic defect of Caucasians (~1/400)
Carrier frequency of 1/10

People absorb & store Fe too efficiently
Dysregulation of intestinal Fe absorption

Symptoms are of Fe toxicity
Oxidative damage to tissues (esp heart and liver failure)
Incr risk of atherosclerosis & CVD

Treat w/phlebotomy, Fe-chelators

Why is this mutation so common?
Normal Hemochromatosis
Liver iron deposition
Genes causing hemochromatosis or
other iron overload conditions
HFE
Surface protein, function unknown
Interacts with TfR
Ferroportin
Alleles with decreased hepcidin response?
Tfr2
Liver specific TfR
Signaling to hepcidin expression?
Hemojuvelin
GPI-linked cell surface protein
Activity affects hepcidin expression
Hepcidin
Ferritin H (IRE mutant)- hyperferritinemia
Ceruloplasmin

JBC 281: 28494 (2006)