HbA NH2

H2O2 Cl2O7
KClO3

NAOH
CH2O
PO4

KMnO4

MEDICINE
KING SAUD UNIVERSITY
COOH
Co2
MgCl2

H2O

SO2
Important Doctors slides
HCN
Extra Information Doctors notes CCl4
CuCl2
SiCl4

Biochemistry Be like stem cells,

Structure and function of differentiate yourself Editing file
from others!
hemoglobin
 By the end of this lecture, the students should
be able to know:
OBJECTIVES
• The structure and function of hemoglobin.

• The factors affecting oxygen binding to hemoglobin.

• Examples of normal and abnormal hemoglobin

structures.
Lecture overview

* Hemoglobin has 2 parts

1- non protein part
2- protein part (heme)
* Myoglobin found in muscles and gives the red color

Recommended video 8:54 mins

Please watch this video before
studying the lecture

*BPG = Bisphosphoglycerate
 Hemoglobin
Hemoglobin (Hb)
– A hemeprotein (heme + protein) found only in red Prosthetic (it is a co factor that is required, when we talk
blood cells. about the enzymes which are not active and required non
– Oxygen transport function. protein part to become active we call it Apo enzyme. And
– Contains heme as prosthetic group. then when the non protein part become attached to it we
Prosthetic means it acts as a co-factor call it holoenzyme so it is not an enzyme it is protein but it
(it doesn’t consume the oxygen itself but it required a non protein part which is heme and it act as
delivers O₂ to the tissues). prosthetic group which is permanently attached to the
– Heme reversibly (not permanent) binds to oxygen. protein
Attached temporarily → co enzyme
Attached permanently → prosthetic group

Enjoy
studying
me
 Hemoglobin
The heme group :
A complex of protoporphyrin IX and ferrous iron (Fe²⁺).
• Fe²⁺ is present in the center of the heme.
• Fe²⁺ binds to four nitrogen atoms of the porphyrin
ring.
• Forms two additional bonds with:
1. Histidine residue of globin chain.
2. Oxygen molecule not atom.

*Heme group has iron in the center in ferrous state , if it is

in ferric state it will not bind to oxygen
*in ferrous state it can have 6 bonds
4 with nitrogen we call it
porphyrin ring
1 bond with molecular O2 (molecule(O2) NOTE atom(O)) The heme group: Fe2+ – porphyrin
1 attached to the globin chain (polypeptide) complex with bound O2
 Hemoglobin
Types of hemoglobin in adults *all the discussion in this lecture will be about hemoglobin A because it
HbA is the majority of the normal adult = 90%
(90%) *HbA1c is a modified form of HbA by adding glucose
HbA2 *chain composition means the types of the globin chains which are
(2%) present
Normal The 4 subunits of the hemoglobin are : 2 alpha and 2 beta and they're
HbF (1%) present as alpha -beta dimers and you can see this in HbA , in HbA1c
the same as HbA but with addition of glucose
HbA1c
(6%) HbF is a fetal hemoglobin it is present in the fetus and by the time of
Hemoglobin
delivery the levels of it goes down so in adult it is less than 2% , it
Carboxy
composed of 2 alpha and 2 gamma chains
Hb
*HbA2 formed before birth and stay for a long time in a very small
Abnormal Met Hb amount

Sulf Hb *Abnormal = unable to carry oxygen

*Carboxy Hb = CO (carbon monoxide) is added NOT CO2
*Met Hb = the iron in ferric state NOT ferrous so can not bind to oxygen
*sulf Hb = amount of sulfa increased in blood because of sulfa
containing drugs or chronic constipation
 HbA
Hemoglobin A (HbA)

– Major Hb in adults.
– Composed of four polypeptide chains:
 Two α two β chains.
– Contains two dimers of αβ subunits (each dimer has 2 heme)
– Held together by non-covalent (hydrophobic) interactions.
– Each chain is a subunit with a heme group in the center that carries oxygen.
– A Hb molecule contains 4 heme groups and carries 4 molecules of O₂

Dimers =(2 sub unites)

*Within the dimer →intradimer bonding it is a strong bond (hydrophobic bonds)
*between 2 dimers→ ionic and hydrogenic bond weaker bond causes a little
movement between 2 dimers

4 subunits binding to 4 hems and each heme can bind to 1 molecule of oxygen so
when it is maximally saturated how many molecules of oxygen can be bind into one
hemoglobin ? Four
 HbA
HbA structure :
T form ( taut form ) R form ( Relaxed form ) When 4 oxygen molecules bind to the
The deoxygenated form The oxygenated form of hemoglobin there will be a
of Hb Hb conformational change in the globin
Taut form Relaxed form chains which will break the ionic bonds
R or relaxed structure of
The movement of dimers The dimers have more oxyhemoglobin = bind to oxygen
is constrained (restricted) freedom of movement =broken ionic bonds
Low-oxygen-affinity form T or taut of deoxyhemoglobin = no
High-oxygen-affinity form
unloaded oxygen oxygen bound=present of ionic bond

‫كيف احفظها ؟‬
‫الريالكسد فورم عندها الكثير من االوكسجين‬
. ‫لذلك تشعر بالحرية وترتبط فيه بشكل قوي‬
‫التوت فورم انسحب منها االوكسجين لذلك‬
‫حركتها صعبة وتكونت عندها عقدة انها ما‬
‫ترتبط باألوكسجين بشكل قوي‬
‫‪HbA:‬‬

‫مجرد صورة توضيحية للجدول السابق‬

 Hemoglobin function and factors affecting O2 binding:
 Hemoglobin functions : about 10% only but the majority of carbon dioxide goes back
 Carries oxygen from the lungs to tissues to the lung as bicarbonate(major buffer) after dissolved in
 Carries carbon dioxide from tissues back to the the blood . Hemoglobin can bind also to protons from
lungs tissues and take them back to the lung
 Normal levels (g/dL):
 Males: 14-16 Factors affecting pH: Negative log of normally (2 molecules of ATP are
O2 binding: ion concentration of formed) 1,3-BPG get converted to
 Females: 13-15
hydrogen. 3-Phosphoglycerate
Inverse relationship But in RBCs glycolysis (no ATP is
Factors that reduce the ability between hydrogen formed) so they take the BPG
of hemoglobin to bind to O2 so ion concentration shunt to convert 1,3-BPG to 3
it can be easily delivered to the and pH value. Phosphoglycerate by 2,3-BPG .
tissues . 2,3-BPG is produced in RBCs and its
3 allosteric effectors: very important abundant there.
When it binds to hemoglobin it
reduces it’s affinity to bind to
pCO2 (partial oxygen which means make the
Po2 (Partial pH of the Availability of 2,3-
carbon dioxide delivery of oxygen to tissues easier
oxygen pressure) environment BPG
pressure)
 Oxygen dissociation curve (the curve is explained in details in the next slide)

The hemoglobin has 4 sites to bind oxygen when the

Oxygen dissociation curve : first molecule binds the globin chain of one subunit it
– The curve is sigmoidal in shape creates a conformational change then the binding of
– Indicates co-operation of subunits in O₂ binding the next molecule will be faster then another
– Binding of O₂ to one heme group increases O₂ conformational change occur and so on, the binding
affinity of others of the last one is the fastest and this is called heme-
– Heme-heme interaction heme interaction .
*myoglobin only one subunit and it can bound to 2
oxygen or don’t bound .

*ODC = Oxygen dissociation curve

Oxygen dissociation curve

In oxygen dissociation curve we measure

the saturation of hemoglobin with oxygen
at different partial pressures of oxygen so
you keep on increasing the amount of
oxygen and notice how the saturation of
hemoglobin is affected .

The hemoglobin has 4 sites to bind

oxygen when the first molecule binds the
globin chain of one subunit it creates a
conformational change then the binding
of the next molecule will be faster then
another conformational change occur and
so on, the binding of the last one is the
fastest and this is called heme-heme
interaction .
 Factors affecting oxygen binding
Factors affecting oxygen binding :

1. pO₂ (partial oxygen pressure) : The mechanism depends on

– P50 (mm Hg): the pressure at which Hb is 50% saturated (Km) concept which is the
with O₂ substrate concentration at
– Indicates affinity of Hb to O₂ : which an enzyme has
A. High affinity → slow unloading of O₂ (low P50 value) achieved half of it is maximal
B. Low affinity → fast unloading of O₂ (high P50 value) velocity which influences the
 Lung pO₂ is 100 mm → Hb saturation 100% affinity . So if the Km value is
 Tissue pO₂ is 40 mm → Hb saturation reduces * high the affinity is low and
 Hence O₂ is delivered to tissues vise versa

*When the oxygen in the lungs :

High partial pressure of oxygen so O2 bind to the Hb , saturated completely and we call it
oxyhemoglobin, 4 oxygen molecules are bound and it goes to tissues
*in the tissues : the oxygen is released from Hb and from the tissues the hemoglobin
carries carbon dioxide and when it binds to hemoglobin we call it carbaminohemoglobin
it goes to the lung
*in the lungs: the CO2 get released from Hb and O2 bind to Hb
*CO2 reduce affinity to O2
When it is released the affinity to O2 increases
 Factors affecting oxygen binding
2. pH of the environment

pH is the negative log of hydrogen ion concentration .

If the hydrogen ion concentration high the ph will be
low and if the hydrogen concentration low the ph will
be high

1- Decrease in pH = increased acidity

2- increasing carbon dioxide pressure
Are both important factors for oxygen delivery
 Factors affecting oxygen binding
3. pCO₂ (partial carbon dioxide pressure) : ‫ هنا‬Bohr effect‫شرحنا لكم‬
– The Bohr effect :
• It is the shift of the ODC to the right in response to an
increase in pCO2 or a decrease in pH The Bohr effect tells you how affinity of Hb to O2
• Is the effect of pH and pCO₂ on: change with changes in amount of the carbon dioxide
A. Oxygenation of Hb in the lungs and (protons =pH) and that is important because you
B. Deoxygenation in tissues are generating carbon dioxide and protons in the
tissue
– Tissues have lower pH (acidic) than lungs *normally without any modifiers or any effectors p50
 Due to proton generation (two reactions): requires 26 mmHg oxygen pressure to achieve 50%
CO₂ + H₂O H2CO₃ saturation pressure
H2CO₃ HCO₃⁻ + H+ *shifting the curve to right = p50 value increased and
– Protons reduce O2 affinity of Hb Causing easier O2 release into that’s mean affinity is decreased (Faster unloading of
the tissues. O2)
– The free Hb binds to two protons. So whenever you need oxygen you will have effectors
– Protons are released and react with HCO3 – to form CO₂ gas to increase p50 values = to shift (OCD) to the right
*shifting the curve to the left = p50 value get reduced
(HCO₃⁻ + H+  CO₂ + H₂O )
– The proton-poor Hb now has greater affinity for O₂ (in lungs). and that’s mean increased affinity and less delivery to
– The Bohr effect removes insoluble CO₂ from blood stream and tissues
produces soluble bicarbonate .
 Factors affecting oxygen binding
4. Availability of 2,3- BPG :
– Binds to deoxy-Hb and stabilizes the T-form.
– When oxygen binds to Hb, BPG is released.

normally (2 molecules of ATP are formed) 1,3-BPG

get converted to 3-Phosphoglycerate
But in RBCs glycolysis (no ATP is formed) so they
take the BPG shunt to convert 1,3-BPG to 3
Phosphoglycerate by 2,3-BPG .
2,3-BPG is produced in RBCs and its very
important abundant there. When it binds to
hemoglobin it reduces it’s affinity to bind to
oxygen which means make the delivery of oxygen
to tissues easier

*When it stabilizes the t form it reduces the affinity to oxygen which means more O2 needs to
be delivered
*due to any reason (e.g. 1- high attitude (hypoxia)
2- anemic people )
the amount of 2,3 BPG increases so it will reduce the affinity of the o2 and more delivery of it
 Factors affecting oxygen binding
Compensatory mechanisms:

At high altitudes and “hypoxia” :

 RBC number increases
 Hb concentration increases
 BPG** increases
 This decreases O2 affinity of Hb
 Thus increases O2 delivery to
tissues

People getting multiple blood transfusions their

2,3 BPG becomes low . When you transfused
blood to a patient , initially the affinity of Hb to
O2 is too high so O2 is not delivered to tissue but
if the patient not very sick (compromised) within
6-24 h they will start form 2,3 BPG
 Factors affecting oxygen binding
Oxygen affinity : Abnormal hemoglobin :
 High O2 affinity is due to: • They are unable to transport O₂ due to abnormal
1. Alkalosis structure
2. High levels of Hb F Sulf-HB: Forms due to Carboxy-Hb: CO₂ Met-Hb: Contains
3. Multiple transfusion of high sulfur levels in replaces O₂ and binds oxidized Fe³⁺
2,3 DPG-depleted blood blood 220X tighter than O₂ (~2%) that cannot
(irreversible reaction) (in smokers) carry O₂

The youtube video in the

first slide explains this
diagram clearly

Recall : Fe²⁺ called Ferrous while Fe³⁺ called Ferric

 Other hemoglobin forms
Fetal Hemoglobin (HbF) HbA2 HbA1c

Composed of two a and two d globin Two α and two β -Glucose

Tetramer with two a and two g chains
chains

Major hemoglobin found in the fetus HbA1c levels are high in patients with
Appears shortly before birth.
and newborn diabetes mellitus
 Higher affinity for O2 than HBA  Constitutes ~2% of total Hb  HbA undergoes non-enzymatic
glycosylation
Important because the maternal O2 Because they have higher amount of  Glycosylation depends on plasma
which is mainly HbA has to be glucose glucose levels
delivered to the fetal hemoglobin *life span of RBCs =120 day so if the
across the placenta which is HbF so glucose bound to HbA1c level
it has to have higher affinity to O2 because it stays for 120 days so until
 Transfers O2 from maternal to fetal
circulation across placenta the patient maintain his blood
glucose level over a period of 3
months
*fasten glucose test will show drop
in glucose because the patient
didn’t eat sugar
 Summary
Hb functions :
Carries oxygen from the lungs to tissues .Carries carbon dioxide from tissues back to the lungs
Normal levels :
Males: 14-16 Females: 13-15
A.High affinity → slow unloading of O₂ (low P50 value)
B. Low affinity → fast unloading of O₂ (high P50 value)
pO₂ - Lung pO₂ is 100 mm → Hb saturation 100%
- Tissue pO₂ is 40 mm → Hb saturation reduces
Acidosis :Decreased O₂ affinity
pH Alkalosis: increased O₂ affinity

The Bohr effect : Is the effect of pH and pCO₂ on

A. Oxygenation of Hb in the lungs B. Deoxygenation in tissues.
Factors affecting oxygen binding

-Tissues have lower pH (acidic) than lungs

CO₂ + H₂O → HCO₃⁻ + H⁺
pCO₂ -Protons reduce O2 affinity of Hb
-The free Hb binds to two protons
- Protons are released (HCO₃⁻ + H⁺ → CO₂ + H₂O )
-The proton-poorHb now has greater affinity for O₂ (in lungs)
-The Bohr effect removes insoluble CO₂ from blood stream and produces soluble bicarbonate .

Availability of 2,3- -Binds to deoxy-Hb and stabilizes the T-form

-When oxygen binds to Hb, BPG is released
bisphosphoglycer At high altitudes and “hypoxia” BPG increases
ate This decreases O2 affinity of Hb , Thus increases O2 delivery to tissues
 QUIZ
Q1 : Which of the following is irreversible ? Q4 : When does HbA2 appears ?
A- Carboxy Hb A- 10 weeks after birth
B- Met Hb B- 12 week after birth
C- Sulf Hb C- 20 weeks after birth
D- Non of the above D- 22 weeks after birth
Q2 : In Met Hb the iron is in the form of … ? Q5 : Which of the following is true about HbA structure ?
A- Ferrous
A- Weak ionic and hydrogen bonds between αβ dimer pairs
B- Ferric
B- Strong interactions between α and β chains from stable αβ
C- Both of them
D- Non of the above
dimer
C- Both of them
Q3 : What is the form in which majority Carbon dioxide is D- Non of the above
transported in the blood ?
A- Bicarbonate Q6 : What do we expect to see in the CBC of a patient from
B- Carbon Monoxide Abha ?
C- Carbonic anhydrase A- High levels of hemoglobin
D- Non of the above B- Low O2 affinity
C- Decreased RBCs
D- Low levels of 2,3 bisphosphoglycerate
 QUIZ
Q7 : Which of the following result from Bohr effect ? Q10 : What are the components of heme group?
A- Removes insoluble CO2 from blood stream
B- Produces soluble bicarbonate A complex of protoporphyrin IX and ferrous iron (Fe²⁺) that binds
C- Produce CO2 to the blood stream to four nitrogen atoms of the porphyrin ring
D- A&B
Q11 : How does oxygen partial pressure affects the oxygen
Q8 : HbA2 is composed of ? binding ?
A- Two α and two γ chains.
B- Two α and two δ globin chains. ↓ p50 ↑ Increase Hb affinity to O2
C- Four α globin chains. ↑ p50 ↓ decrease Hb affinity to O2
D- Two α and two β chains.
Q12 : What are normal levels of hemoglobin ?
Q9 : determine three factors that induces high O₂
affinity ? Males: 14-16
Females: 13-15
1. Alkalosis
2. High levels of Hb F
3. Multiple transfusion of 2,3 DPG-depleted blood
Suggestions and recommendations

1) C 2) B 3) A 4) B 5) C 6) A 7) D 8) B
 TEAM
MEMBERS

TEAM LEADERS
Saleh Altwaijri
Mohammad Almutlaq Leen Altamimi
Rania Alessa Laila Mathkour

Muneerah Alzayed
THANK • Lippincott's Illusrated Reviews Biochemistry 6 th E

YOU
FOR CHECKING 10:06 mins

OUR WORK
Review the notes

PLEASE CONTACT
US IF YOU HAVE @436Biochemteam
ANY ISSUE

Biochemistryteam436@gmail.com