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HOW PROTEIN
LEARNING OUTCOMES
STRUCTURE 1

 At the end of the lecture, students should be

DETERMINES able to:

 describe the properties of structural protein
such as collagen and globular protein such
FUNCTION as haemoglobin.
 relate the importance of sequence of amino
DR. KHAIZURIN TAJUL ARIFIN acids and its organization into 3-
DEPARTMENT OF BIOCHEMISTRY dimensional structure to give rise to
biological function.
 illustrate the detrimental effects of gene
mutation which leads to abnormal function
of protein.
 explain the relationship between protein
and its function in health and disease.

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FIBROUS PROTEIN: FIBROUS PROTEIN:

COLLAGEN COLLAGEN
• Produced mainly by • 3 polypeptide chains : H bonds
• fibroblasts, • Every thirds amino acid is Gly
• muscle cells and • Collagen: (Gly-X-Y) repeats
• epithelial cells • Y: frequently Pro or hydroxyproline
• Type I / collagen(I) is the most abundant in mammals • X: any other aa
• Precursor: procollagen(I) • What is hydroxyproline?

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FIBROUS PROTEIN: COLLAGEN

• Hydroxyproline:
H bond
• Hydroxylysine:
sites of
attachment for
disaccharides

Fig 49.3. Hydroxylation of proline and lysine residues in collagen. Proline and lysine residues within the
collagen chains are hydroxylated by reactions that require vitamin C and Fe2+

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COLLAGEN DISEASES:
COLLAGENOPATHIES OTHER FIBROUS PROTEINS
• Ehlers-Danlos Syndrome ELASTIN
– mutations in the gene for
type III collagen-deficiency • Major protein in elastic fibres
of lysyl hydroxylase or • Who / which structure needs to be elastic?
procollagen peptidase or
mutations in the amino acid • Located in ECM of connective tissue
Stretchy skin of Ehlers-Danlos sequences-col type III- • Precursor: tropoelastin
syndrome vascular problems.
• Osteogenesis Imperfecta • Rich in Pro and Lys, a little of hydroxyPro and
– type I collagen, known as
hydroxyLys
brittle bone syndrome.
• Marfan’s Syndrome
– mutations in the fibrillin gene

OI

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OTHER FIBROUS PROTEINS OTHER FIBROUS PROTEINS

• Role of 1-antitrypsin (A1AT) in elastin
degradation LAMININ
• Met residue in A1AT important for • Most abundant protein in basal laminae
binding
• Binds to type IV collagen and other molecules in the
• A1AT deficiency→emphysema
ECM

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PROT STR DETERMINE FUNCTION:

HEMOGLOBINS
GLOBULAR HEMEPROTEINS

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HEMOGLOBIN A STRUCTURE 13 14

Consists of 4 subunits (or polypeptide chains)

Each subunit has similar structure as myoglobin
Its oxygen-binding properties are regulated by interaction with allosteric
effectors
HbA
Identical dimers= (αβ)1, (αβ)2
Hydrophobic interaction within the dimer αβ
Ionic and hydrogen bonds hold the dimers (weaker)

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OXYGEN BINDING EQUILIBRIA FOR BINDING OF

OXYGEN MOLECULES TO HB
• O2 binds directly to the Fe2+
• Hb exists in 2 alternate conformation
• The pulling of O2 binding moves the
• T (tense) state (low affinity for O2, binding
helix
sites are hindered)
• Function of myoglobin is • R (relaxed) state (higher affinity
unaffected
• Positive cooperativity
• Loss of salt bridges →
conformational changes
• Cooperativity of O2 binding in Hb

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Oxygen binding and Heme

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OXYGEN DISSOCIATION 18

CURVE

Oxygen dissociation curve

Fe2+ in the centre is held by bonds to the 4 nitrogens with 2 additional bonds-side chain of a
histidine residue and oxygen on each side of the planar porphyrin ring.

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ALLOSTERIC EFFECTS
ALLOSTERIC EFFECTS
1. Haem-haem interactions 2. Bohr effect
a) Loading and unloading oxygen • ↓ pH (acidic or basic?) or
b) Significance of sigmoidal oxygen
 pCO2,
dissociation curve?

Release of O2 from Hb   
Oxygen affinity of Hb  or ↓?

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ALLOSTERIC EFFECTS ALLOSTERIC EFFECTS

2. Bohr effect 2. Bohr effect
a. Where do the protons come from? b. Mechanism of Bohr effect
- Metabolically active tissues

- In tissues: carbonic anhydrase

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ALLOSTERIC EFFECTS ALLOSTERIC EFFECTS

3. Effect of 2,3-BPG on oxygen affinity 3. Effect of 2,3-BPG on oxygen affinity
- [2,3-BPG] ~ [haemoglobin] b. Binding site of 2,3-BPG
a. Binding of 2,3-BPG to deoxyhaemoglobin - 2,3-BPG expelled on
- decreases oxygen affinity of Hb oxygenation of Hb

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ALLOSTERIC EFFECTS ALLOSTERIC EFFECTS

3. Effect of 2,3-BPG on oxygen affinity 3. Effect of 2,3-BPG on oxygen affinity
c. Shift of the oxygen dissociation d. Response of 2,3-BPG levels to chronic
hypoxida or anaemia
curve
- [2,3-BPG]  in response to chronic
- 2,3-BPG expelled on oxygenation hypoxia
of Hb
e. Role of 2,3-BPG in transfused blood
- Stored blood: decreased 2,3-BPG and
abnormally
high oxygen affinity
- Transfused RBC able to restore 2,3-BPG
6-24h

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TRANSPORT OF CO2
ALLOSTERIC EFFECTS
4. Binding of CO2
- Most CO2 produced in metabolism is transported as
bicarbonate ion
- Some as carbamate – N ter hameoglobin –
carbaminohaemoglobin
- Stabilizes deoxy form – what happens to oxygen affinity?
What happens to oxygen dissociation curve?

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ALLOSTERIC EFFECTS
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5. Binding of CO
PH (BOHR EFFECT)
-CO binds tightly to Hb – H+ bind to Hb —>
carboxyhemoglobin reduce Hb affinity for
-220x affinity to Hb O2
HbO2 + H+ → HbH +
O2

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EFFECT OF GENE MUTATIONS SICKLE CELL ANEMIA (HbS)

 2s2=HbS
 A point mutation in the
gene for  chains.
• Primary structure  results in production of
– Alterations in DNA sequence (mutations) mutant hemoglobin.
alter the amino acid sequence  The polar amino acid
glutamic acid at position
– Alter the folding and function of protein six has been replaced
– Example: sickle cell anaemia with nonpolar amino
acid valine.
 An infant- does not
showing symptoms of
the disease. Why?

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SICKLE CELL ANEMIA (HBS) SICKLE CELL ANEMIA (HBS)

• Characterized by A to T
• lifelong episodes of pain (crises)
• Chronic hemolytic anemia with
associated hyperbilirubinaemia
• Increased susceptibility to infections
in early childhood

• Caused by
• Point mutation in DNA (A to T
substitution)
• Leads to anoxia
• Cause pain, eventually death of cells
• Decreased oxygen tension, increased pCO2, decreased pH, dehydration,
increased 2,3-BPG in erythrocytes increase sickling

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HBS OTHER
HEMOGLOBINOPATHIES

• Generating a hydrophobic sticky patch (protrusion) • Hemoglobin C disease

– the surface of the subunit of deoxyHbS • Hemoglobin SC
– fits into a complementary site on the alpha chain of
another Hb molecule. • Methemoglobinemias
• Low pO2 deoxy HbS can polymerize inside the RBC • Thalassemias **
forming a gel
– assembling into a network of fibrous polymer that stiffen
and distort
– producing rigid erythrocytes.
• Sickled cells frequently block the flow of blood in the
narrow capillaries.
• Caused localised anoxia-pain-death of cells.

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THALASSEMIAS
• Imbalance in synthesis of globin chains
• -Thalassemia
• -Thalassemia?

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