Blood

Magdalena Mielczarek-Puta PhD

Chair and Departament of Biochemistry
 The blood
• a type of connective, liquid tissue circulating in the blood vessels system

• consist of liquid plasma (a cellular substance) and cells

(morphotic elements)

• an adult human contains 4-6 liters of blood (about 75 ml / kg)

• men usually have 1 liter of blood more than women

CONSTITUENTS OF BLOOD
WHOLE BLOOD
Serum
blood plasma without fibrinogen and other clotting factors
COMPONENT OF BLOOD PLASMA
 PLASMA PROTEIN
• total plasma protein 6 - 8g/dl
- albumins 3.5 – 5 g/dl
- globulins 2.5 – 3.5 g/dl
- fibrinogen 200 – 400 mg/dl

• all plasma proteins except gamma protein are synthesized in liver

• gamma fraction – produced by plasma cells located in the lymph node

A/G ratio 1.2-1.5 to 1

higher solubility

lower solubility
 ALBUMINS
• the major constituent of blood plasma protein (3.5 – 5 g/dl)
• 25% of total hepatic protein synthesis
• half life 20 days

• molecular weight – 66,5 kDa

STRUCTURE

Teritary structure 17 disulfide bonds

ALBUMINS
MAINTENANCE OF COLLOIDAL OSMOTIC PRESSURE
 EDEMA
• Low albumin level (below 2 g/dl called hypoalbuminemia) leads to edema

Edema – palpable swelling caused by an incerase in interstitial fluid volume

Causes of hypoalbuminemia

Excessive loss of protein Insufficient synthesis

• nephrotic syndrome – • liver failure – cirrhosis, hepatitis
- lost protein in urine - kidney damage • low protein diet
• hemorrhage • incomplete protein in diet
• burns
• exudative gastroenteropathy
 TRANSPORT FUNCTION OF ALBUMIN

Albumin as high hydrophilic protein bind various hydrphobic molecules

Copper
BINDING SITES IN ALBUMIN
GLOBULINS
(glycoprotein)
NORMAL PATTERN OF PLASMA PROTEIN ELECTROPHORESIS
ABNORMAL PATTERN OF PLASMA PROTEIN ELECTROPHORESIS

multiple myeloma
 HEMOGLOBIN STRUCTURE

ferrous Fe(II)

HbA - a tetrameter composed of two identical α chains (141 aa residues)

and two identical ß chains (146 aa residues)

each chain contains one heme molecule with Fe2 + atom (binds oxygen) in the center
 HEME STRUCTURE
• The prostetic group consists of
- an iron atom (Fe II) in the center
linked to N in pyrrole rings with differ in substituents
• Four pyrrole rings linked together by a methene bridge
• Heme is non-covalent bound to proximal histidine (F8) of globulin
 Hemoglobin - conformational changes
Quaternary structure –
two dimers (αß) 1 and (αß) 2 connected with each other by weak ion and hydrogen
bonds.

Between the α and ß chains in each dimer there are strong hydrophobic bonds

Hemoglobin exists in two distinct states: the T-state and the R-state.
The T-state "Taut" - deoxyhemoglobin".
The R-state "Relaxed" - oxyhemoglobin (fully oxygenated form)
 Hemoglobin – oxygen binding

Binding one oxygen molecule to Fe changes the structure of tetramer

Oxygen moves Fe into the plane of the pyrrole rings what attracted proxymal
histidine

Displacement of neighboring amino acids leads to breaking of cross-links between

the carboxyl residues of all four globin subunit what increases affinity for oxygen
 Cooperative oxygen binding

Binding 1 oxygen molecule to hemoglobin

causes an increase in oxygen affinity
of the remaining subunits
(affinity to 4th molecule of oxygen is 300 times
higher than to the first)

Release of one oxygen molecule from hemoglobin

causes a decrease in the affinity
Oxygen dissociation curve
 DISTAL HISTIDINE - FUNCTION

After binding oxygen ferrous undergoes oxygenation (Fe II) not oxidation (Fe III)

• The distal histidine (E7) in Hb strongly conserved AA

• A hydrogen bond between the N epsilon proton of E7 and the second oxygen atom
stabilize O2 bound to the ferrous iron
 CARBON MONOXIDE POISONING

• Carbon Monoxide (CO) - a dangerous gas (odorless and colorless)

• Sources of CO - running automobiles and gas-powered appliances
• When inhaled, it binds at the same sites as oxygen and negatively impact
the body's ability to absorb oxygen
 ANOTHER SIGNIFICANCE OF DISTAL HISTIDINE
• CO binds to hemoglobin 200 times more tightly than oxygen
• Even at low partial pressures, CO will prevent Hb from delivering O2 to the body
• Once CO binds to one site of hemoglobin, hemoglobin turns into the R-state
which increases oxygen affinity and prevents oxygen dissociation in tissues
• Treatment of CO poisoning - the administration of 100% oxygen at higher partial
pressures (displace most of CO from Hb)
Affnity of free hem
to CO is 25000 times
higher than O2
TYPE OF HEMOGLOBIN
 ABNORMAL HEMOGLOBIN

• hemoglobinopathies - substitution of one amino acid for another

- the synthesis of abnormal hemoglobin eg. HbS, HbC, HBSC

• thalasemia alfa or beta - inherited disorders

– lack of alfa or beta chain synthesis
- the severity thalassemia depends on how many genes are missing
 Polymerization of HbS

 

  
  
  
   Association is repeated

   over and over to

 produce large, rod-like

aggregates that bind

 oxygen poorly and

distort shape of
erythrocytes
34
 ALPHA THALASSEMIA

The degree of impairment is based on which clinical phenotype is present

(how many genes are affected)
BETA THALASSEMIA

Excess of HBF and HBA2

 HEMOGLOBIN FUNCTION

• transport of gases (oxygen, carbon dioxide)

• buffering function
BINDING OF OXYGEN IN THE LUNGS
 RELEASE OF OXYGEN IN THE TISSUES

• only 25% of Hb (one monomer) releases O2 in resting tissues

• O2 is released first from beta than from alfa chain
 CARBONIC ANHYDRASE

• present in erythrocytes, kidney and stomach

• one of the fastest enzyme
• catalyzes reversible reaction

pH decrease
bind to Hb

diffuse to blood plasma

BOHR EFFECT
BOHR EFFECT
 2,3 BISPHOSPHOGLYCERATE IN ERYTHROCYTES

• product of lateral glycolysis pathway

• important in erythrocytes
• bypassing substrat-level phosphorylation
 EFFECT OF 2,3 BPG ON OXYGEN AFFINITY

2,3 BPG binds only to beta chains therefore does not affect fetal hemoglobin HbF
HbF vs HbA

• HbF carries 20-30% more oxygen than HbA

• gamma subunits have higher affinity for
oxygen than beta subunits
• 2,3 BPG does not affect HbF
because is bound only to beta subunits

HbA
TRANSPORT OF OXYGEN AND CARBON DIOXIDE - GAS EXCHANGE
TRANSPORT CARBON DIOXIDE
HEME SYNTHESIS
HEME SYNTHESIS
 PORPHYRIAS

• Porphyrins accumulation - negatively affecting the skin or nervous system

• The types that affect the nervous system - acute porphyria

• Symptoms are rapid in onset and last a short time

• Symptoms include abdominal pain, chest pain, vomiting, confusion, fever,

high bood pressure

• Complications may include paralysis, low blood sodium level, seizuria

• Attacks - triggered by alcohol, smoking, drugs (metabolized in cyt. 450)

or hormonal changes, fasting, stress

• The types that the skin is affected - hepatoerythropoietic porphyria

• Symptoms include blister or itching and occur after sunlight exposure

porphyrynogens accumulation – after UV light – reactive forms – destroy skin
 PORPHYRIAS – „VAMPIRE DISEASE”

Legends about vampire - person suffering from porhyria

drink blood, avoid sun lights, have blister and ichting on skin
avoid garlic – metabolized by cyt.450 (contains heme)

History: Notable cases

Vincent van Gogh
King George III „Maria the Mad” Lead poisoning
HEME CATABOLISM
 TYPE OF BILIRUBIN

Unconjugated bilirubin Conjugated bilirubin

Called - bound with albumin Called – bilirubin diglucuronide
- indirect bilirubin - direct bilirubin

UDP-glucuronyl
transferase

Bloodstream
Liver
BILIRUNIN METABOLISM
 JAUNDICE

also known as icterus, is a yellowish or greenish pigmentation of the skin

and whites of the eyes due to high bilirubin levels (hyperbiliruminemia)

is clinically obvious when plasma bilirubin concentration exceed 3 mg/dL

PREHEPATIC (HEMOLYTIC) JAUNDICE
 NEONATAL JAUNDICE

Type of physiological, hemolytic jaundice

Reasons:
• exchange HbF into HbA
• immature coniugated system in liver
(immature UDP-glucuronyl transferase,
lack of glucuronic acid)
 HEPATIC JAUNDICE

urobilinogen

The causes:
Infection (vital hepatitis)
Toxic chemicals (alcohol, chloroform)
Drugs liver disorder
Cirrhosis
POSTHEPATIC JAUNDICE

, gallstones

gallstones in gallbladder
JAUNDICE SUMMARY
Changes in pH below 6.8 and above 8 may result in death
 pH OF BLOOD
bicarbonate buffer - the largest buffering capacity (75%)

carbonic anhydrase

Henderson–Hasselbalch equation

20
pH = 6.1 + log 1

pH = 6.1 + 1.3
pH = 7.4
ACIDOSIS AND ALKALOSIS

20
pH = 6.1 + log 1

pH = 6.1 + 1.3
pH = 7.4
ACIDOSIS AND ALKALOSIS – compensatory mechanism
 ACIDOSIS AND ALKALOSIS

20
pH = 6.1 + log 1

pH = 6.1 + 1.3
pH = 7.4
carbonic anhydrase
pentose
ATP
ADP

ATP
ADP
 ERYTHROCYTES METABOLISM
Anaerobic glycolysis (lack of mitochondrion)
• transformation of glucose to lactate
• produces only 2 ATP molecules per glucose molecule

- 1 ATP

- 1 ATP

+ 2 ATP

NAD regeneration
by lactate dehydrogenase

+ 2 ATP

2 ATP
PENTOSE PHOSPHATE PATHWAY IN RBC - significance
THE END
THANK YOU