Chapter 11

Blood Biochemistry
 Teaching target
1. Master the basic raw materials of synthetic heme, the key enzymes
and the basic process.
2. Familiar with heme synthesis regulation.
• Composition of the blood
– The liquid element: plasma
– The formed elements: red blood cells
(erythrocytes), white blood cells
(leukocytes), and platelets
(thrombocytes).
• Plasma --- The blood fraction obtained after
removal of the cellular components.
• Serum --- The liquid element obtained by
allowing the blood to clot.
 Blood is a fluid tissue composed
of plasma and blood cells that are
circulating in cardiovascular system.
Blood volume accounts for 7%～8 % of body weight
 Functions of Blood
1. Play roles in maintaining homeostasis
(1) Buffer (pH of the blood)
(2) Transport ( gases, nutrients, waste products,
hormones, etc)
(3) Regulate （body temperature）

2. Participate in defense function

(1) Defense reactions
( humoral and cell-mediated immunity)
ˊ
(2) Hemostasis
I. Composition
1. Plasma (50% ~ 55% of blood volume)
2. Blood Cells

(WBC)

( RBC )
I. Composition
1. Plasma ( ~ 55% of blood volume)
① H2O ( 91% ~ 92% )
② Plasma proteins ( 6.5% ~ 8.5% ) (Functions-
p68)
globulin
albumin colloid substance
clotting factors (colloid osmotic pressure)
③ Electrolytes
Na+ K+ crystal substance

Ca2+ Cl- （crystal osmotic pressure）

④ Others
glucose, amino acid, lipids, vitamins, gases,
hormone, end-products, transmitters etc.
I. Composition
1. Plasma
2. Blood Cells
① RBC (erythrocyte): 120d
② WBC (leukocyte): 3~4d
③ Platelet : 7~14d
( active in first 2 days )
I. Composition
1. Plasma
2. Blood Cells
Hematocrit (Ht )

percentage of blood
volume occupied by
all blood cells

40% ~ 50% in men

37% ~ 48% in women
(45 %)
Standard Blood Smear
RBC and Stained WBCs
Permeability
Osmosis
Red Blood Cell (Erythrocytes)
• Structure
– Biconcave,
anucleate
– Plastic deformation
• Components
– Hemoglobin
– Lipids, ATP,
– Carbonic anhydrase
• Function
– Transport O2 from
lungs to tissues
and CO2 from
tissues to lungs
Hemoglobin
 Erythropoietin

• Production of red blood cells

– Stem cells proerythroblasts early erythroblasts
intermediate late reticulocytes
• Erythropoietin: Hormone to stimulate production of RBC
 Leukocyte (WBC)
Granulocyte (contain many
“granules”,organelles)
– Neutrophile
– Eosinophile
– Basophile
Agranulocyte
– Lymphocyte Morphology →Histology
– Monocyte Functions →Immunology
Medicine
 Platelet

It is not a cell.
Cytoplasmic fragments of megakaryocytes.
It is filled with granules involved in blood clotting.
(serotonin, Ca2+, ADP etc.)
 Platelet Adhesion

血小板与非血小板表面的粘着
Platelet Release Reaction
Platelet Aggregation
Hemostasis
Pathway of blood coagulation
Plasma proteins
 Plasma proteins

 concentration 65 –80 g  l
 simple or conjugated (glycoproteins, lipoproteins)
 separation:
 a) salting-out methods  albumin, globulins,

fibrinogen
 b) electrophoresis  albumin, globulin 1, 2, , 

fractions: - 


2 1
+
 Elfo fractions of plasma proteins

Fraction Rel. amount c (g/l)

(%)
Albumins: albumin 52 – 58 34 – 50
pre-albumin (transthyretin)
1-globulins: thyroxin-binding globulin, transcortin, 2,4 – 4,4 2-4
1-acid glycoprotein, 1-antitrypsin, 1-lipoprotein
(HDL), 1-fetoprotein

2-globulins: haptoglobin, macroglobulin, ceruloplasmin 6,1 – 10,1 5–9

-globulins: transferrin, hemopexin, lipoprotein (LDL), 8,5 – 14,5 6 – 11

fibrinogen, C-reactive protein, C3 and C4 components of
the complement system

-globulins: IgG, IgM, IgA, IgD, IgE 10 – 21 8 – 15

 Plasma proteins participate in:

1. blood coagulation
2. maintenance of homeostasis (pH, osmotic pressure)
3. defence against infection
4. transport of
nutrients
metabolites
hormones metabolic waste
drugs
 General properties of plasma proteins

 Most are synthesized in the liver

Exception: -globulins – synthesized in plasma cells

 Synthesized as pre-proteins on membrane-bound polyribosomes;

then they are subjected to posttranslational modifications in ER
and Golgi apparatus

 Almost all of them are glycoproteins

Exception: albumin

 They have characteristic half-life in the circulation (albumin – 20

days)

 Many of them exhibit polymorphism (immunoglobulins,
transferrin…)
 Acute phase reactants (APRs)

 Their levels change during acute inflammatory response

 APRs concentration changes in:

 infection

 surgery

 injury

 cancer
Acute phase reactant response
Types of APRs:

Positive

1-antitrypsin
Negative
C-reactive
protein (CRP): albumin
~1000-fold
transferrin
increase!
fibrinogen
haptoglobin (HP)
 ALBUMIN

 Concentration in plasma: 45 gl

  60% of the total plasma protein

 Functions:
 maintenance of the osmotic pressure of plasma

 transport of:

• steroid hormones
• free fatty acids
• bilirubin
• drugs (sulfonamides, aspirin)
• Ca2+
• Cu2+
 Causes of Albumin Deficiency

 Liver diseases (cirrhosis) – decrease in the ratio of

albumin to globulins

 Protein malnutrition

 Excessive excretion by kidneys (renal disease)

 Mutation causing analbuminemia (affects splicing)

Blood: Cells and Plasma Proteins
I. Major functions of blood and its components:
 transport and distribution of essential nutrients to
tissues
 removal of waste products
 defense against internal insult
 repair of damaged tissues

II. Two major components of blood: liquid phase and formed

elements
Plasma - blood treated with anticoagulants to prevent clot
formation then centrifuged to remove the cells

Plasma
Plasma
“Buffy
Buffycoat”
coat
Red blood cells
Red blood cells

Serum - blood that is allowed to form a fibrin clot then

centrifuged to remove the cells and the clot
Red Blood Cells
(Erythrocytes)

 1011 produced per day

 Anucleate

 Lack intracellular organelles (incapable of

protein synthesis and repair)

 Contain specific proteins and ions

 Transport oxygen, and remove CO2 and H+

 Finite life span of 60-120 days before

being destroyed in the spleen
Platelets
 2-3 mm in diameter

 150-400 x 106/mL blood

 Normal subcellular structures except they

lack a nucleus

 Capable of mitochondrial protein synthesis

 Platelet function is dependent upon their

activation

 Involved in blood clotting

 Contain secretory organelles called alpha-

granules, which contain proteins important in
blood clotting

 Circulate for 10 days before being destroyed

 Plasma proteins

Synthesized by liver or produced by plasma (B) cells

A. Albumin
B. Transferrin Transport proteins
C. Ceruloplasmin
D. Enzymes – coagulation enzymes, complement factors
E. C-reactive protein – acute phase reactant
F. Immunoglobulins – humoral immunity
 ALBUMIN

66 kDa

Accounts for ~50% of the total plasma protein and ~ 50%

of the total liver protein production

 Half-life ~ 20 days

 Highly polar

 At pH 7.4 it is anionic with 20 negative charges per molecule

 Functions of Albumin

 Transport of long chain fatty

acids & sterols
 Regulation of colloidal pressure

Decreases in albumin concentration

cause edema

Gaw: Clinical Biochemistry; Churchill Livingstone (1999), p. 44.

 Causes of decreased plasma albumin:
I. Decreased synthesis
A. malnutrtion
B. malabsorption
C. advanced chronic liver disease

II. Abnormal distribution or dilution

A. overhydration
B. increased capillary permeability like in
septicemia

III. Abnormal excretion or degradation

A. nephrotic syndrome
B. burns
C. hemorrhage
D. certain catabolic states
E. protein losing enteropathies

IV. Rare congenital defects

A. hypoalbuminemia
B. analbuminemia
 TRANSFERRIN
 Transports iron in plasma as ferric
ions (Fe3+) – each transferrin molecule
binds 2 Fe3+

 Protects the body against the toxic

effects of free iron

Normally 30% saturated with Fe3+ -

increased or decreased saturation is
indicative of iron overload or deficiency,
respectively
 CERULOPLASMIN

Major copper transport protein

 Regulates oxidation-reduction,
transport and utilization of iron

 Increased concentrations are

found in active liver disease or
tissue damage

 Decreases are sometimes

observed in individuals with
Wilson’s disease
 ENZYMES
 Enzymes of the coagulation and complement cascade have
a defined function in blood

 Others reside elsewhere in the body and appear

incidentally in the blood – their measurement is of
diagnostic value

Alanine
Creatine kinase
aminotransferase

Acid phosphatase Amylase

 C-REACTIVE PROTEIN (CRP)

 Major component of the acute phase response

and a marker of bacterial infection

 Mediates the binding of foreign polysaccharides,

phospholipids and complex polyanions, as well as
the activation of complement

 <1 mg/mL in normal plasma

 Slightly elevated levels of CRP are indicative of

chronic, low-grade inflammation and have been
correlated with an increased risk of
cardiovascular disease
 IMMUNOGLOBULINS

 Proteins produced and secreted

by B cells specific for the foreign
particle that stimulated their
synthesis

 Five classes of Ig

 Multiple myeloma: arises from

the aberrant proliferation of a
single B cell. Light chains of Igs
are produced in excess of heavy
chains = paraproteins.

Elevated serum protein levels

are usually due to an increase in
the Ig fraction and may indicate
the presence of a paraprotein.
§1 Heme Synthesis
Sites:

Mitochondria and cytoplasm

Precursors:

Glycine, succinyl CoA, Fe2+

1. Synthetic pathway of heme
(1) The formation of -aminolevulinic acid (ALA)

COOH COOH

H2C CH2NH2 HSCoA + CO2 H 2C

CH2 + CH2
COOH ALA synthase
C～SCoA (pyridoxal phosphate) C O

O CH2NH2
In the mitochondria
(2) The formation of porphobilinogen (PBG)

COOH
O
CH2
OH
CH2
HO
O C
ALA O
H C H dehydratase

H N H
2H2O N
H2N H

In the cytosol
(3) The formation of uroporphyrinogen Ⅲ
(UPGⅢ) and coproporphyrinogen Ⅲ
(CPGⅢ)
Deaminase Linear
4× Porphobilinogen tetrapyrrol

UPG Ⅲ
isomerase
UPGⅢ
decarboxylase
CPGⅢ UPGⅢ

In the cytosol
(4) The formation of heme

CPGⅢ
oxidase
CPGⅢ Protoporphyrinogen Ⅸ
Protopor-
phyrinogen Ⅸ
oxidase
ferrochelatase
Heme Protoporphyrin Ⅸ

In the mitochondria
 PBG
Heme

Gly
succinyl CoA Linear
Protoporphyrin Ⅸ tetrapyrrol

Protoporphyrinogen
Ⅸ
CPGⅢ UPGⅢ
2. Regulation of heme biosynthesis

(1) ALA synthase

• Rate-limiting enzyme
• Feedback-inhibition by heme
• Intensity inhibition by Ferriheme
• Inducing by some drugs and
hormones (eg. testosterone)
(2) ALA dehydrase and ferrochelatase
Inhibited by heme, and heavy metals.

(3) Erythropoietin (EPO) can increase

synthesis of heme and hemoglobins.
BLOOD DISORDERS
ANEMIA- a condition in which you don't have enough
healthy red blood cells to carry adequate oxygen to
your tissues
 Iron Deficiency Anemia
Inadequate amounts of iron to form hemoglobin in
the red blood cells

Causes
• insufficient iron in the diet
• poor absorption of iron by the body
• ongoing blood loss, most commonly from menstruation or
from gradual blood loss in the intestinal tract
• periods of rapid growth
TREATMENT
• Iron Supplements
• Increasing iron in your diet (green leefy vegetables)
 Aplastic Anemia

• Aplastic Anemia – bone marrow does not produce enough

RBC
• Causes:
• Chemotherapy and radiation
• toxic chemicals
• Virus
• Some rheumatoid arthritis medications and even
some medications
• Idiopathic
 Pernicious anemia
• A decrease in red blood cells that occurs
when the intestines cannot properly absorb
vitamin B12.
• Causes
• A special protein, called intrinsic factor
(IF), helps your intestines absorb vitamin
B12. This protein is released by cells in the
stomach. When the stomach does not make
enough intrinsic factor, the intestine cannot
properly absorb vitamin B12.
 Aplastic Anemia

• TX:
• Blood transfusions
• Eliminating the cause
• Bone marrow transplant

• Frequently FATAL!
 Pernicious Anemia
• Treatment
• Vitamin B12 injections once a month
• Vitamin B12 absorbed in the mouth
• Vitamin B12 given through the nose
 Hemorrhagic Anemia
• A type of anemia caused from heavy blood loss
due to bleeding somewhere in the body.
Causes
• Large blood loss in life-threatening situation such
as traumatic injury, massive gastrointestinal
hemorrhage, such as ulcers, hemorrhoids, gastritis
(inflammation of the stomach) and cancer.
Treatment
• stop bleeding (both internally and externally)
• IV saline, plasma or albumins
• Possible blood transfusions for large amounts of
blood loss.
 Leukemia
• Type of cancer
• Overproduction of immature white
blood cells
• They take the place of RBCs

• Treatable with bone marrow

transplants, chemothemotherapy,
radiation
Blood Smear of a patient
with Leukemia

Normal blood smear

Blood Smear; Leukemia
 St. Jude Hospital

Leukemia is one of the most common childhood cancers. It

occurs when large numbers of abnormal white blood cells fill the
bone marrow and sometimes enter the bloodstream.

Because these abnormal blood cells are defective, they don't help
protect the body against infection the way normal white blood
cells do. And because they grow uncontrollably, they take over
the bone marrow and interfere with the body's production of
other important types of cells in the bloodstream, like red blood
cells (which carry oxygen) and platelets (which help blood to
clot).
 Infectious mononucleosis
sometimes called "mono"
or "the kissing disease," is
an infection usually caused
by the Epstein-Barr virus
(EBV).

EBV is very common, and

many people have been
exposed to the virus at
some time in childhood.

What white blood cell do

you think would be elevated
 Blood poisoning - Septicemia
• An infection enters the blood stream
• Can be deadly
• Treated with antibiotics (usually IV
antibiotics to start)
 SICKLE CELL ANEMIA
• Genetic Disorder
• Abnormally shaped blood
cells
• Parents can be carriers
(asymptomatic)
• Common in African
American, Hispanics an
and Mediterranean
dissent
Sickle Cell Anemia is actually
codominant

AA = normal
Aa = sickle cell trait (few symptoms)
aa = sickle cell anemia
If both parents are
carriers, child has a ¼
chance of having the
disease
Complications

1. Pain
• Lethargy
• Lifelong anemia
(low red blood count)
• Organ failure
• Stroke
• Jaundice (yellowing of the skin
and whites of the eyes). It occurs
because the rapid breakdown of
abnormal red blood cells leads to
a build-up of a waste product in
the body called bilirubin
 Treatments
• There is no cure for Sickle cell
• Treatment is aimed at relieving pain,
preventing infections and organ
damage.
• Some may be cured with bone marrow
and stem cell transplants but it is risky
and more research is being done.
 HEMOPHILIA
This disorder causes a failure of the
blood to clot
Patients can be treated with blood
transfusions that include clotting agents.
Hemophilia is carried on the X
chromosome

Females X H X H normal
X H X h carrier
X h X h hemophiliac

Males X H Y normal
X h Y hemophiliac
 Research
• Research what clotting factors are missing with the
different forms of hemophilia.
• Symptoms
• Treatments
• Prognosis
• What diseases could they be susceptible too and
why?
• What is thrombocytopenia?
• What vitamin may doctors prescribe before surgery to
help our blood clot? Why?
• What is a normal WBC count and What WBC Counts
do we see with leukemia?
 Methemoglobinemia
• a disorder
characterized by
the presence of a
higher than normal
level of
methemoglobin
(metHb) in the
blood
• Methemoglobin is an
oxidized form of
hemoglobin that has
a decreased affinity
for oxygen, resulting
in an increased
affinity of oxygen to
other heme sites
and overall reduced
ability to release
oxygen to tissues
• The oxygen–
hemoglobin
dissociation curve
is therefore shifted
to the left. When
methemoglobin
concentration is
elevated in red
blood cells, tissue
hypoxia can occur.
• Normal methemoglobin levels are <1%
• Elevated levels of methemoglobin in the blood are
caused when the mechanisms that defend against
oxidative stress within the red blood cell are
overwhelmed and the oxygen carrying ferrous ion
(Fe2+) of the heme group of the hemoglobin
molecule is oxidized to the ferric state (Fe3+).
• This converts hemoglobin to methemoglobin,
resulting in a reduced ability to release oxygen to
tissues and thereby hypoxia.
• This can give the blood a bluish or chocolate-
brown color.
 2 types
• Congenital
– Genetic defect present at birth (recessive gene)
• Aquired
– Exposure to exogenous oxidizing drugs and their
metabolites (such as benzocaine, dapsone and
nitrates) may accelerate the rate of formation of
methemoglobin up to one-thousandfold,
overwhelming the protective enzyme systems and
acutely increasing methemoglobin levels.
– Infants under 6 months of age are particularly
susceptible to methemoglobinemia caused by nitrates
ingested in drinking water (called blue-baby syndrome
 Treatment
• Methemoglobinemia can be treated with
supplemental oxygen and methylene
blue1% solution 1 to 2 mg/kg administered
intravenously slowly over five minutes
followed by IV flush with normal saline.
Methylene blue restores the iron in
hemoglobin to its normal (reduced)
oxygen-carrying state.
PROGRESS
Puping Liang, Chenhui Ding,
Hongwei Sun et al. Correction of
β-thalassemia mutant by base
editor in human embryos.
Protein & Cell, First Online: 23
September 2017,
doi:10.1007/s13238-017-0475-6
 Questions
1. What are the common characteristics of
plasma proteins?
2. Describes the function of plasma proteins.
3. What are the characteristics of the synthesis of
heme?