Immunoglobulin: Structure and Function

By Markos Negash (Bsc,Msc)

 CHAPTER 8

Immunoglobulin: Structure and Function

 Learning Objectives
Up on completion of this chapter the student will be
able to know :
• About Introduction to Immunoglobulins and
antibodies
• General Functions of Immunoglobulin
• Basic immunoglobulin Structure
• Immunoglobulin Fragments: Structure/Function
• Human immunoglobulin classes
 Antibodies
• Antibodies(Abs) are serum glycoproteins
synthesized by plasma cells

• Collectively, Abs form a family of glycoproteins

known as Immunoglobulins

• Abs specifically bind antigens in both the

recognition phase and the effector phase of
humoral immunity
• Abs are distributed in biologic fluids
throughout the body and are found on the
surface of a limited number of cell types
 BASIC STRUCTURE OF IMMUNOGLOBULINS
• All Ab molecules share the same basic structural
cxs but display remarkable variability in the
regions that bind Ag
A. Heavy and Light Chains
• All Igb have a four chain structure as their basic
unit

• This are two identical light chains (23Kd) and

two identical heavy chains (50-70Kd)
B. Disulfide bonds
• The heavy and light chains and the two heavy
chains are held together by

• inter-chain disulfide bonds and

• by non-covalent interactions

• The number of interchain disulfide bonds

varies among different Igb molecules.
• C. Variable (V) and Constant (C) Regions

• Both the heavy and light chain could be divided into

two regions based on variability in the aa sequences

1. Light Chain -VL (110 aa) and CL (110 aa)

2. Heavy Chain -VH (110 aa) and CH (330-440 aa)

• variable region, composed of 110-130 aa, give the

Ab its specificity for binding Ag
D. Hinge Region -The region at which the arms
of the antibody molecule forms “Y” is called
the hinge region

• because there is some flexibility in the

molecule at this point.
E. Domains - 3D images of the Igb shows that it
is not straight

• Rather, it is folded into globular regions each

of which contains an intra-chain disulfide
bond

These regions are called domains.

• 1. Light Chain Domains -VL and CL
• 2. Heavy Chain Domains -VH, CH1 -CH3 (or
CH4)
• H chains of IgM and IgE have 5 domains (VH,
CH1, CH2, CH3 and CH4). There is no hinge
region in these two isotypes

• F. Oligosaccharides – CHO are attached to the

CH2 domain in most Igb

• However, in some cases CHO may also be

attached at other locations
BASIC STRUCTURE OF IMMUNOGLOBULINS
 STRUCTURE OF THE VARIABLE REGION

• A. Hypervariable (HVR) or complementarity

determining regions (CDR)
• Comparisons of the amino acid sequences of
the variable regions of Ig's show that most of
the variability resides in three regions called
the hypervariable regions or the
complementarity determining regions.
• Ab with different specificities (i.e. different
combining sites) have different CDR's

• While Ab of the exact same specificity have

identical CDR's (i.e. CDR --> Ab Combing
site)

• CDR's are found in both the H and the L

chains.
• B. Framework regions
• The regions between the CDR's in the variable
region are called the framework regions (FR)

• Based on similarities and differences in the

framework regions the Igb HC and LC
variable regions divided into groups and
subgroups.

• These represent the products of different

variable region genes.
 Immunoglobulin Fragments: Structure/Function
Relationships

• Igb fragments is produced by proteolytic

digestion

• Fab: Digestion with papain breaks the Igb

molecule in the hinge region before the H-H
inter-chain disulfide bond

• This results in the formation of two identical

fragments that contain the LC and the VH and
CH1 domains of the HC
•Fc - Digestion with papain also
produces a fragment that two HC
each containing a CH2 and CH3 • P
a
domain p
a
i
n
•This fragment was called Fc
because it was easily crystallized.

•The Fc region of the molecule is

digested into small peptides by
pepsin. • F
• F c
a
• F(ab')2 -Treatment of Igb with pepsin results
in cleavage of the HC after the H-H inter-chain
disulfide bonds

• This fragment is called F(ab')2 because it is

divalent

• The F(ab')2 binds antigen but it does not

mediate the effector functions of antibodies.
 Effector function of Antibodies

• Many of the effector functions of Ab are mediated

by the HC constant regions of Igb molecules

• different Ig heavy chain isotypes serve distinct

effector functions

• all these functions are triggered by the binding of

antigens to the variable regions
 Effector function
1.Neutralization of microbes and microbial toxin
-Ab against microbes and microbial toxins block
the binding of these microbes and toxins to
cellular receptors
2.Antibody mediated opsonization and
phagocytosis

• Abs of the IgG isotype coat (opsonize) microbes

by bindings to Fc receptors

• Leukocyte Fc receptors promote the phagocytosis

of opsonized particles and deliver signals that
stimulate the microbicidal activities of the
leukocytes
 3.Antibody-Dependent cell –Mediated
Cytotoxicity

• Fc receptor of NK cells called FcyRlll (CD16),

binds to IgG Ab attached to cells, and the result is
lysis of the Ab-coated cells

• This process is called Ab-Dependent cell-Mediated

Cytotoxicity (ADCC)
-Engagement of FcyRlll by Ab coated target cell
activates the NK cells
-To synthesize and secrete cytokines such as
IFN γ
-Discharge the contents of their granules
4. Complement activation
 Immunoglobulin Classes, Subclasses, Types And
Subtypes
• The Igb can be divided into 5 different classes
based on differences in the aa sequences in the
constant region of the HC

1. IgG -Gamma (γ) heavy chains

2. IgM -Micro (μ) heavy chains
3. IgA -Alpha (α) heavy chains
4. IgD -Delta (δ) heavy chains
5. IgE -Epsilon (ε) heavy chains
Immunoglobulin Subclasses
• The classes of Igb can de divided into
subclasses based on small differences in the aa
sequences in the CR of the HC

• 1. IgG Subclasses
a) IgG1 -Gamma 1 (γ1) heavy chains
b) IgG2 -Gamma 2 (γ2) heavy chains
c) IgG3 -Gamma 3 (γ3) heavy chains
d) IgG4 -Gamma 4 (γ4) heavy chains
• 2. IgA Subclasses
a) IgA1 -Alpha 1 (α1) heavy chains
b) IgA2 -Alpha 2 (α2) heavy chains
• Immunoglobulins can also be classified by
differences in the LC

• differences are detected by serological means.

1. Kappa light chains
2. Lambda light chains
• The LC can also be divided into subtypes
based on differences in the aa sequences in the
CR .
1. Lambda subtypes
a) Lambda 1
b) Lambda 2
c) Lambda 3
d) Lambda 4
Genetic Variation in Antibody Heterogeneity
• Isotypes
• Allotypes
• Idiotypes

ISOTYPES
• Definition - Isotypes are antigenic determinants
that characterize classes and subclasses of HC
chains and types and subtypes of LC
• Location –HC isotypes are found on the Fc portion
of the CR of the molecule while LC isotypes are
found in the CR

II. ALLOTYPES
• Definition -Allotypes are Ag determinants specified
by allelic forms of the Igb genes

• Allotypes represent slight differences in the aa

sequences in the HC or LC of different individuals
• Even a single aa difference can give rise to an
allotypic determinant, although in many cases the
several aa substitutions have occurred

III. Idiotypes
• Definition -Unique Ag determinants present on
individual Ab molecules or on molecules of
identical specificity

• Identical specificity means that all Ab molecules

have the exact same HVR
Source: Kuby. Immunology 2007 5 th ed).
 Structure And Some Properties Of Ig Classes
And Subclasses
IgG
Structure:
• All IgG's are monomers
• -The subclasses differ in the number of
disulfide bonds and length of the hinge region.
Properties
1.Most versatile Igb because it is capable of carrying
out all of the functions of Igb molecules

2.IgG is the major Igb in serum (75%) and in extra

vascular spaces

3.Placental transfer - IgG is the only class of Igb that

crosses the placenta via receptor on placental cells
for the Fc region
• Not all subclasses cross equally; IgG2 does not cross well

4.Fixes complement - Not all subclasses fix equally well;

IgG4 does not fix complement

5. Binding to cells-Macrophages, monocytes, PMN's and

some lymphocytes have Fc receptors for the Fc region of
IgG

• Not all subclasses bind equally well; IgG2 and IgG4 do

not bind to Fc receptors
• Binding of IgG to Fc receptors on other types
of cells results in the activation of other
functions
• A consequence of binding to the Fc receptors
on those cell can now internalize the antigen
better.
• The term opsonin is used to describe
substances that enhance phagocytosis. IgG is a
good opsonin.
 IgM
Structure
• IgM normally exists as a pentamer but it can
also exist as a monome

• In the pentameric form all heavy chains are

identical the valence is theoretically 10

• IgM has an extra domain on the H chain

(CH4) and it has another protein covalently
bound via a S-S bond called the J chain
•This chain functions in polymerization of the molecule
into a pentamer

J Chain

Cµ4
• Properties
1. IgM is the 3rd most common serum Ig

2. IgM is the first Ig to be made by the fetus and

the first Ig to be made by a virgin B cells when it
is stimulated by Ag

3. As a consequence of its pentameric structure,

IgM is a good complement fixing Igb
4.IgM is also a good agglutinating Ig . Thus, IgM
Ab are very good in clumping microorganisms
for eventual elimination from the body

5. IgM binds to some cells via Fc receptors

6.B cell surface Ig -Surface IgM exists as a

monomer and lacks J chain
 IgA
• Structure
• Serum IgA is a monomer but IgA found in
secretions is a dimer

• When IgA exits as a dimer, a J chain is

associated with it

• When IgA is found in secretions is also has

another protein associated with it called the
secretory piece or T piece
• The secretory piece made by epithelial cells, helps
IgA to be
–transported across mucosa and
– protects it from degradation in the
secretions
• Properties
1.IgA is the 2nd most common serum Ig

2. IgA is the major class of Ig in secretions -

tears, saliva, colostrum, mucus. Since it is
found in secretions secretory IgA is important
in local (mucosal) immunity

3. Normally IgA does not fix complement,

unless aggregated.
 IgD

• IgD exists only as a monomer.

• Properties
1. IgD is found in low levels in serum; its role in
serum uncertain.
2. IgD is primarily found on B cell surfaces
where it functions as a receptor for antigen.
• IgD on the surface of B cells has extra amino
acids at C-terminal end for anchoring to the
membrane.
3. IgD does not bind complement.
 IgE

• Structure-IgE exists as a monomer and has an

extra domain in the constant region.
Properties
1. IgE is the least common serum Ig since it binds
very tightly to Fc receptors on basophils and
mast cells even before interacting with antigen

2. Involved in allergic reactions

3. IgE does not fix complement.

4. plays a role in parasitic helminth diseases. Since
• serum IgE levels rise in parasitic diseases,

• Eosinophils have Fc receptors for IgE and

binding of eosinophils to IgE-coated helminths
results in killing of the parasite.
THANK YOU!!!