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- Iron Biological Role

Tuesday, January 15, 2019 1:33 PM

LS3N03_201
9_M1W1_Iron
Hazardous rad
macromolecule
 Iron easily oxidizes
- Ferrous - reduced

Highly abundant but because it is easily oxidize in aerobic

environment
- It precipitates and becomes an issue because cells can
use it - to the ferric state
○ Easily goes from ferrous to ferric or back based
condition
- Cells needs to ensure that you cannot leave free iron i
places because it becomes unavailable
- It has to be bound to a protein
- Cannot have iron free in the cell
- Iron highly reactive and can be detrimental to cells
- Participates in phantom reactions
○ In ferrous state
○ Forms free radicals
§ Damage the DNA
□ Oxidizes it or breaks it
§ Disrupt membrane morphology
□ Damages phospholipids
□ Any membrane of organelle
§ Alter cell-signalling
□ Change oxidation state of proteins o
proteins themselves
dicals that can attack § Oxidizes macromolecules that are needed
les and cause tissue injury the structure and function of the cell
- Cells need to get rid of free iron in cell to minimize
radicals
n't

on

in

or

d for
Hazardous rad
macromolecule
dicals that can attack §
les and cause tissue injury the structure and function of the cell
- Cells need to get rid of free iron in cell to minimize
radicals

Cell minimizes free iron in cell by using

- Proteins that bind to iron to avoid it from being free
○ They proteins also use iron to function properly

3 main groups:
- Prosthetic groups
○ Ex. Heme group
- Sulfur cluster
○ Organic component + iron coordinated by differe
atoms
§ Dependent on the number of atoms
- AA side chains
○ Of a protein
○ See examples on slide
○ Transferrin - important
§ Protein that binds iron by certain AA that
coordinate iron
□ Carbonate bond
ent
Quite a bit of variation in proteins that bind iron

Nitrogen atoms help coordinate Fe in heme

- Heme = iron-protoporphyrin 9
 That contain or bind iron

- In blood

- Store in muscle

- Generate ATP
Nitrogen atoms help coordinate Fe in heme
- Heme = iron-protoporphyrin 9
- Heme B most common

Difference between Hemo and Myo

- Structural different
○ Heme - 4 subunits
○ Myo - 1 subunits
§ Heme is stable by hydrophobic interactions
with AA that face the pocket
§

 (oxygen-carrying po
hemoglobin and my

- Erythrocytes
 ○ Myo - 1 subunits
§ Heme is stable by hydrophobic interactions
with AA that face the pocket
§

ortion of the
yoglobin)

Hemoglobin
- 4 subunits
- There are infant hemoglobin and adult hemoglobin
- Adult
- 2 alpha
- 2 beta
- Coordinates FERROUS species of iron
- In order to carry oxygen, it must be in the FERROU
state
- Ferrous can make 6 bonds
§ 4 - nitrogen to stabilize molecule


US
 - 2 beta
- Coordinates FERROUS species of iron
- In order to carry oxygen, it must be in the FERROU
state
- Ferrous can make 6 bonds
§ 4 - nitrogen to stabilize molecule
§ 1 - oxygen
§ 1 - polypeptide chain
- Each hemoglobin carries 4 oxygen because there are 4
heme groups in hemoglobin
- Binding of oxygen occur due to oxygen in the ferrous st
in the heme group

Hemoglobin has a high infinity for CO than O

- Due to the way it binds to the heme group
○ Specifically the iron
- If you have same molecules of CO than O, more CO will
bound to hemoglobin
○ Level of CO cause asphyxiation
US

tate

be
 Prefer

Before the release of the oxygen, the electron shift reverses

Equilibrium - only difference is the state of iron

- Darker in colour - brown

○ Diagnosis of high
○ Normal - up to 1%
- Cannot carry O2 or CO2
erential binding of CO to hemoglobin -> Asphyxiation

Oxidation of the iron in the hemoglobin (one extra electron)

- Can generate free radical
- But because it is contained in the hemoglobin, then it
stays in and doesn't affect cell
○ Neutral to cell
- Must be in ferrous state to bind iron

If you have hemoglobin without oxygen

- And exposed to chemicals as oxidants
- Oxidants may oxidize iron in hemoglobin
- Makes ferric iron in the hemoglobin
- This is METHEMOGLOBIN
○ Hemoglobin with ferric iron

n
level of methemoglobin
%, any more is detrimental
2
- Darker in colour - brown
○ Diagnosis of high
○ Normal - up to 1%
- Cannot carry O2 or CO2
- Causes cyanosis and hyp

Uses reduction

Non-enzymatic

Non-enzymatic
 n
level of methemoglobin
%, any more is detrimental
2
poxia

Cell (RBC) has mechanisms to keep it down at 1%

- Two mechanisms:
1. Inactivate oxidants in cell
- Different ways
□ Reducing the oxidant
® Non-enzymatic ways
n more - more efficient ® Like reaction between oxidant
substances like ascorbic acid
® Not enough oxidant to oxidize
hemoglobin
□ Enzymatic
® Inactivate oxidants present in t
cell
◊ Ex. Catalyses or peroxida
Enzymatic ® Forms water and oxygen
® Reducing the oxidant
2. Reduce level of methemoglobin - bigger roles
<- most effective - Reduce any methemoglobin that has forme
- Different ways
□ Non-enzymatic ways
□ Enzymatic
® NADH-MET - best way of
reduction
® If you have problem with the
enzymes, then other pathways
t and

the

the

ases

ed

s
 Non-enzymatic

- Me

<- standa
- As
- No
- Me
No treatment
 □ Non-enzymatic ways
□ Enzymatic
® NADH-MET - best way of
reduction
® If you have problem with the
enzymes, then other pathways
would be better

Enzymatic

Acquired
- OD on drugs
- Preservatives
○ Nitride based
ethemoglobin [MET Hb] in RBC > 1-2%
Hereditary - rare
- Due to mutation with coding sequence of
hemoglobin or subunits
○ Hemoglobin M instead of B
○ Heterozygous
○ 25-30% of MetHb
§ No cure!
- Due to mutation or defiency in the NADH-
enzyme that breaks down methemoglobin
○ Can be homo or heterozygous

ard treatment = ascorbic acid or IV methylene blue

scorbic acid puts the iron back into the ferrous state
on-enzymatic way of reducing methemoglobin
ethylene blue speeds up the other ways to reduce methemoglobin

RBC - good way of recycling iron in the body

- Major source in body
- More than from intake
s

-MetHb
n
No treatment
 RBC - good way of recycling iron in the body
- Major source in body
- More than from intake
- Just a little more in food to make up for what is
feces

In RBC
- Part of maturation
- Major source of iron in body by recycling old dying RBC
- Each contains millions of hemoglobin
- 2/3 of iron in body is in circulating blood in the RBC
○ The rest in the liver or organ

Heme synthesis
- Heme = prosthetic group containing iron as cofactor
- Transports oxygen
- In RBC
s lost in

Don't need to know how the subunits are formed - know there are 2 groups built separatel

2 enzymes:

By endocytosis

If levels low, iron may

come from storage protein
within cells - ferritin
- All done at the same
time
ly that come together
Synthesis
- Rise from precursor
- Heme group has to have an iron
- Iron comes from extracellular space
○ Endocytosis of proteins that bind iron
§ Transferrin TF

Tf - transferrin
- Main carrier of iron in blood
Holo-Tf - transferrin with 2 irons ions (in ferric state - 3+) boun
- The more iron in the blood, the more iron will be bound
transferrin - up to 2
- May just bind 1 if the level of iron is low
TfR1 - transferrin receptors of red blood cells
- Binds holo-Tf

Once inside cells in endosomes, there are different mechanism

iron from endosome to the mitochondria
- Metformin - gets the iron out
- Multistep process with many reactions
○ Some in mitochondria, some in cytoplasm
- Synthesis of heme group goes back and fourth

Protoporphyrin - everything except for the iron

Iron initially from recycling of dying RBC eaten by macrophage

intestines

FC - adds iron on porphyrin ring

Synthesis of heme group and different subunits (alpha and bet

simultaneously
- All one multi-step process

Some steps in mitochondria, some in the cytoplasm

nd
d to

ms to get

es and from

ta)
Synthesis of heme group and different subunits (alpha and bet
simultaneously
- All one multi-step process

Some steps in mitochondria, some in the cytoplasm

Cells recycle transferrin into plasma to pick up more iron and u

ALAS2 - one of the initial enzymes needed at the beginning

Having iron or synthesizing iron is the rate-determining step

- Anemia when there are low levels of iron
○ No need to synthesis the alpha and beta subunits
no iron
○ Cell can sense the amount of iron in the blood and
determine how much hemoglobin is needed to be
synthesized

Mitoferrin - regulates the amount of iron from cytoplasm to th

mitochondria

Heme groups are the largest groups that use iron as a cofactor
- But there are other proteins with different functions
ta)

used again

if there is

d
e

he

r
 <- main tr

<- used by tissues to s

ransporter of iron in blood

store iron

Can bind to iron in FERRIC state

- Released by liver
- Synthesized by hepatocytes
○ One cell type in the liver
- Apo-tf - when no iron bound
- Can bind up to 2 atoms of iron
○ In ferric state
- Holo-tf - when it has 2 irons
- Has a physiology pH and state
- Saturation = 30%
○ Means you have a lot of transferrin in blood t
have iron bound to it
○ To ensure that there is a lot of transferrin read
away iron that is free in cytoplasm or in plasm
§ It can generate radicals
§ And precipitate - not available anymore
□ Essential but useless when precip
○ Body avoids having irons free in the blood
§ High level of transferrin and many with
bound to it
○ Can bind when there is an iron overdose
§ Up to a certain level

pH changes the conformation of a protein and changes th

for the substrate or cofactor
that doesn't

dy to take
ma

e - useless
pitated

no iron

he affinity
 bound to it
○ Can bind when there is an iron overdose
§ Up to a certain level

pH changes the conformation of a protein and changes th

for the substrate or cofactor

<- one carbonate that assists with binding

Main storage protein to store iron

- Because having free iron is bad
- In case there is low iron levels
- Used by all cells
- Up to 24 subunits
○ Heavy and light
- Different variations of ferritin have different ratios

Ferritin - has iron in soluble state ready to use

Hemosiderin - insoluble making iron less accessible to the cell
- Aggregates of ferritin
- When there is too much iron in the blood
- To extra store iron in an insoluble way

Endosome have lower pH than cytoplasm

Iron comes in at ferrous state but stored as ferric iron

- Gets oxidized when it is stored in the core of the protein

Pay attention to redox state of iron

- Because proteins are picky
- Some bind ferrous and some bind to ferric

Ferritin used to synthesize heme group

Any cells can store iron as ferritin and use when needed
- Especially cells that are dividing
he affinity

n
Ratio H/L is cell type- and iron loading-dependent
Any cells can store iron as ferritin and use when needed
- Especially cells that are dividing

24 in total but ratio change dependent on the cell type

- Cell can control the synthesize of the subunit based o
of iron at a certain time

H-rich
Have H to L ration
- Rich in H subunit
- Less capable of storing iron
- But iron is more accessible
- Easier to release

L-rich
- More L than H
- Has more iron but less accessible
- Accumulate when too much ferritins around
- Live and spleen

Expression on the synthesis of subunit dependent on the ir

environment

If there is too much iron in the cytoplasm

- Make more L-rich subunits

Holo-tf interact with two receptor

on the load

ron in the
Tfr1 - most cell
Tfr2 - liver and intestine
<- might be
 Holo-tf interact with two receptor
- R1 - higher infinity
- R2 - lower infinity

As the endosome forms, it is also up taking the DMT1

- It becomes part of the endosome membrane

Iron inside the lumen - making the pH more acidic

- Causing release of iron
e needed as a cofactor - But in ferric state
- So it needs to be oxidized inside the lumen
- Since the DMT1 can transport in the ferrous state

Get familiar with the name of the receptors and cofactors

FERRIC STATE
 Don't wa

<- easily accessible

<- because its not free in the blood or cytoplasm

<- how many transferrin receptors are available

<- how many iron bound to those t

- How much iron is available
 Iron is a cofactor

ant it free in the cytoplasm or blood

Also protect organism from infections

- Even bacteria
- Ensuring that there is no free iron makes it not available from pathogens

e in the cell - the most expressed, the more iron can be uptake

transferring

There is no such thing as free iron in the cell

- Because ferritin would absorb it