Small letters indicate proteolytic fragments: a = smallest fragment b = largest fragment

Fig 2.20

C3b is an opsonin recognized by complement receptor 1 (CR1)

Thioester

Fig 2.28

 Functions of C5a (C3a helps):

C3a

Activates phagocytosis of C3b oposonized pathogens bound to Complement Receptor 1 (see Fig 2.27) Causes chemotaxis of macrophages and neutrophils through C5a receptor With C3a: Causes increased permeability of blood vessels Fig 2.25 right side

Lectin Pathway
Mannan binding lectin and Ficolins associate with MBL associated serine proteases: MASP 1 & MASP 2 Upon binding of the lectin to mannose or other carbohydrates the MASPs activate
Can now cleave C4 and C2

Thioester

Fig 2.16

Fig 2.31

Antigens, Antibodies & T-cell receptor structure

Chapter 4

Antibodies
AKA: immunoglobulins

Produced by B cells
Can be located:
On the surface of B-cells (B-cell receptor) Secreted forms found in fluids throughout body: plasma, mucous & intercellular fluids (antibody)

Antibodies II
Two main functions:
1. Bind specifically to molecules of the pathogen

2. Recruit other immune system cells and molecules to destroy the pathogen

Five classes: IgM, IgA, IgD, IgG, IgE

Antibodies II
Two main functions:
1. Bind specifically to molecules of the pathogen

2. Recruit other immune system cells and molecules to destroy the pathogen

Five classes: IgM, IgA, IgD, IgG, IgE

MADGE

Antibody structure
Basic unit has Y structure
B-cell receptor has addition of transmembrane domain

4 polypeptides: 2 light (L) chains (27 kD) and 2 heavy (H) chains (55 or 70 kD)
Pairs are identical in a single immunoglobulin molecule

One light chain attached to each heavy chain Heavy chains bound to each other

Disulfide bonds hold it together

Fig 4.2

Antibody structure III

Heavy & light chains both contain N-terminal variable (V) region & C-terminal constant (C) region Heavy chain: 1 V domain (VH), 3 or 4 C domains (CH)

Light chain: 1 V domain (VL), 1 C domain (CL)

Heavy and light V regions combine to form antigen binding site
(2 per immunoglobulin molecule)
Fig 4.1

Antibody structure IV
C regions interact with effector proteins & cells Separation of recognition & effector parts by a flexible hinge region (Fig 4.3) Two parts can be separated proteolytically giving Fab and Fc

Domain structure
Each is composed of a similar folded structure: immunoglobulin fold Disulfide bond between cysteines holds together sheets made of 7 or 9 antiparallel beta strands

Main feature of Immunoglobulin (Ig) superfamily

V Domain structure (Fig 4.7)

Most differences between V domains of different Ig molecules are confined to three stretches of ~10 aa: hypervariable regions These are connected by non-variable framework regions 3 VH and 3VL hypervariable regions fold together to form antigen binding site (Fig 4.7) Also called complementary determining regions (CDRs)

Fig 4.7 Note: CDR3 most variable and forms most extensive contacts with antigen, but CDR1 and CDR2 plus some framework residues are also involved
Fig 4.7

C domain structure
Differences in CH structure used to define: Antibody Classes (Isotypes):
IgM, IgA, IgD, IgG, IgE

Antibody Subclasses:
IgA1, IgA2 IgG1, IgG2, IgG3, IgG4

C domains of all members of each isotype or subclass have essentially the same amino acid sequence

C Domain structure II
Heavy chain C domain gene nomenclature uses Greek letter for appropriate isotype: IgG1 = Cg1, IgD = Cd IgM & IgE heavy chain C regions: 4 Ig

IgA, IgD & IgG heavy chain C regions: 3 Ig

Membrane bound forms have a hydrophobic C-terminus

Fig from last edition of book

Effector functions
A different C region means a different effector function Mediated by proteins that bind to constant domains (eg: Fc receptors on macrophage)

The isotype of a specific antibody can change during a response (IgM to IgG)
Isotype switching: Change in the type of CH domain associated with the same specific V domain

Light chain isotypes

k and l: distinguished by C regions Antibodies have 2 k or 2 l, but never one of each Differ in amino acid sequence but no known functional differences

A typical human has 60% k and 40% l containing antibodies

Antigen binding
Antigen: substance bound by antibody or T-cell receptor

Immunogen: molecules that stimulate immune responses

It is possible for small molecules to be antigens but not be immunogens

Hapten: a small molecule linked to a carrier to make it immunogenic Polyvalent or multivalent antigens have multiple repeated binding sites for antibodies: epitopes

Epitopes
The structure or shape recognized by an antibody (Fig 4.8)

Types of Epitopes
Linear epitope: formed by several adjacent amino acids (approximately 6)
Conformational epitope: formed by amino acids that are not in a linear sequence, but are close together in the folded structure

Neoantigenic epitope: created by modification of the covalent structure (e.g. phosphorylation)

Antigen binding
Antigen is bound through non-covalent interactions
hydrogen bonds, electrostatic, Van der Waals forces, hydrophobic forces (Fig 3.9)

Affinity: Strength of binding between a single antigen binding site and an antigen Kd: dissociation constant - concentration of antigen that binds 50% of the binding sites (e.g. 10-9 to 10-11 M)
Note the smaller this number the stronger the binding

Avidity: overall strength of attachment based on number of binding sites

T Cell Receptor
Ig superfamily member receptor Restricted to CD4+ and CD8+ T lymphocytes Antigen binding domain contains 3 CDRs
CDR3 is the main antigen contact CDR1 and CDR2 contact Major Histocompatibility Complex (MHC) receptors on another cell

TCR:MHC affinity lower than Antigen: antibody interactions (Kd = 10-5 to 10-7)

What did we learn today?

Structure of antibodies
5 isotypes (MADGE) Constant and variable domains Immunoglobulin fold CDR variability

Antigen binding
Epitopes