Enclonar, Kimberly / MLS 3A

Complement Proteins
January 26, 2021
Lean Kristin Ugdang, RMT

Complement proteins
• Series of more than 30 proteins that amplify the
immune response and destroy and clear foreign
substances
o Only a few participate in activate; the rest
participate in inhibition
• Complement function of antibodies and phagocytic
cells
• Activated in a CASCADE manner and results in the
formation of membrane attack complex (MAC) that
lyse the foreign cells Classical Pathway
• Some complement proteins and its products of • Both IgM and IgG are capable of fixing and activating
activation acts as opsonin and anaphylatoxins complement in classical pathway

Activation Pathways Recognition Unit

• Classical Pathway • Target cells must be opsonized first by IgM or IgG to
o Primarily involves 9 proteins form antigen-antibody complex
o Triggered by ANTIGEN-ANTIBODY complexes • Activation unit: C1
o Antibodies involved: IgG ang IgM o Globular structure with 3 subunits (C1q, C1r,
• Alternative Pathway C1s) stabilized by calcium
o Originally called "properdin pathway" ▪ C1q - 6 strands that forms 6 globular heads
o Triggered by LPS, fungal cell walls, yeast cells, ▪ C1r - 2 subunits
viruses, virally infected cells and tumor cells ▪ C1s - 2 subunits
o These sites are binding sites for the opsonin C3b • C1 molecules bind to antibodies rather than antigens
• Lectin Pathway • Binds to Fc region; at least 2 heads of C1q should bind
o Activated by mannose or similar sugars to antibodies for activation
o IgG - CH2 domain
o IgM - CH3 domain
Basic Antibody Structure
• Monomeric - one globular molecule • Binding of C1q to CH domains of Fc region sends a
o 2 Fab - 2 binding sites mechanical stress to the entire C1 molecule
o 1 Fc region - effector region; interacts with stimulating activation of serine protease C1r →
immune cells • Activation of C1s which has C4 and C2 → marks the
• Has 4 to 5 domains end of recognition stage
o 1 in variable region (VH)
o 3-4 in constant region (CH1; CH2; CH3; CH4) Activation Unit
• IgM • Begins from the cleavage of C4 and ends with the
o Pentamer (5 monomeric units) production of the C5 convertase
o 10 Fab, 5 Fc • C1s → cleaves C4 into C4a & C4b → C4a circulation to
o 5 domains act as anaphylatoxins → C4b either binds to surface
o Adopts a staple form microbe or to water molecules resulting in
• IgG inactivation
o Monomer • C1s → cleaves C2 → C2a + C2b →
o 2 Fab, 1 Fc o C2b does not have enzymatic activity therefore
o 4 domains will not continue
o C2a binds to C4b → C4b2a (C3 convertase) - key
• Globular Y shape - typical shape of antibody
to amplification of pathway
• C3 - major complement component of serum with
1,200ug/L
• Cleave of C3 = pivotal points for all pathways
• 1 C3 convertase can cleave 200 C3 (large amounts of
C3a and C3b)
o C3a - does not proceed, anaphylatoxin
o C3b
▪ Bind to c4b2a → c4b2a3b (classical
pathway c5 convertase) - end of activation
unit
▪ Go to site of injury and opsonized
organisms (potent opsonin)
 Enclonar, Kimberly / MLS 3A
Lectin Pathway
• Complement activation in the innate immune
response in the absence of antibodies
• Non-specific recognition of CHO that are commonly
shared by microbes
• Mannose binding Lectin (MBL) - calcium dependent
o Opsonin that binds to mannose found on cell
surfaces of bacteria, viruses, yeast and some
parasites
o Structure:
▪ Structurally similar to C1
▪ 1 subunit of MBL, with 6 globular heads
(carbohydrate recognition domain) - same
with C1q
▪ 2 subunits of MASP-1 (mannose-associated
serine protease 1) - same with C1r
▪ 2 subunits of MASP-2 - same with C1s
o Requires calcium as cofactor
• Activation is similar with classical pathway
o MBL binds to the surface of microbes
recognizing the presence of mannose and similar
sugars
o This binding triggers a conformational change of
the MBL → auto-activation of MASP1 →
activates MASP2 → activates C4 and C2

Membrane Attack Complex (MAC)

• Pore forming complex that creates holes on the cell
membrane of the microbial cell
• C5b attaches to cell membrane of microorganisms,
forming the beginning of MAC
• C5b + C6 → C5b6 + 1 C7 (hydrophobic site of C7 Alternative Pathway
inserts to the lipid bilayer of membrane) → c5b67 + • Amplification loop of the complement activation since
C8 (forms small hole; slow lysis can be observed; lysis it activates in between the classical and lectin
of RBCs but not nucleated cells) → C8 induces pathways
polymerization of 10-16 molecules of C9 (complete • Only needs C3b opsonization to the microbial surfaces
MAC) to activate
• ~100 A - channel • C3b production
o Classical or lectin must be activated to produce
• In cell membrane - hydrophobic
• In external channel - hydrophilic C3b
o Water and enzymes in plasma can also hydrolyze
• Disruption of membrane leads to
o Loss of cell homeostasis C3 to C3a and C3b but at slow rate - only small
o Disruption of protein gradient amounts of C3b is produced
o Penetration of hydrolytic enzymes (lysozymes)] • Does not require antibody opsonin
o Death • Can play in the early innate defense
• Properdin pathway - properdin was thought to play a
major role
• Factors affecting activation
o Bacterial cell wall (LPS, fungal cell wall, virally
infected cells, parasites eg Trepanosomes)
• Steps
o C3b binds to bacterial cell wall + factor B → C3bB
o Binding of factor B makes it more susceptible to
cleavage by factor D (serine protease)
o Factor D cleaves factor B to Ba and Bb → Bb
remains forming C3bBb (alternative pathway C3
convertase) → unstable + properdin = C3bBbP
o C3bBbP → C3a (anaphylatoxin) + C3b
o C3b
▪ Bind to alternative C3 convertase forming
C3bBb3b (alternative pathway c5
convertase)
 Enclonar, Kimberly / MLS 3A
▪ Others act as opsonins and activate more
pathways
o C5 convertase initiates formation of membrane
attack complex by cleaving to C5a and C5b

Anaphylatoxins
• C3a, C4a, C5a
• Help promote and modulate inflammation by
inducing smooth muscle contraction
• Activate mast cells and basophils to release histamine
(vasodilator)
• Vasodilation
• Mediate chemotaxis, inflammation, and generation of
cytotoxic oxygen radicals.
• C5a - most potent