Autoimmune Diseases: Experimental Therapy

with Protein A. ELECTIVE

IMMUNOSUPPRESSION.
Dmitri Popov. PhD, Radiobiology. MD
(Russia)
Advanced Medical Technology and
Systems Inc. Canada.

Protein A
Protein Ais a 42kDasurfaceprotein

originally found in the cell wall of the

bacteriumStaphylococcus aureus. It is
encoded by the spagene and its regulation
is controlled by DNA topology, cellular
osmolarity, and atwo-component system
called ArlS-ArlR. It has found use in
biochemical research because of its ability
to bindimmunoglobulins.

Protein A.

Protein A.
It is composed of five homologous Ig-

binding domains that fold into a three-helix

bundle. Each domain is able to bind
proteins from many mammalian species,
most notablyIgGs. It binds the heavy chain
within theFc regionof most
immunoglobulins and also within theFab
regionin the case of the human VH3
family. Through these interactions in serum,
where IgG molecules are bound in the
wrong orientation (in relation to normal
antibodyfunction), the bacteria disrupts
opsonizationandphagocytosis.

Protein A.
Purified (unconjugated) Thermo Scientific Pierce

Recombinant Protein A is useful as the basis for preparing

various kinds of probes or affinity media for detection or
purification of rabbit and human antibodies, especially IgG
isotypes, in immunoassays and antibody purification
protocols.
Features of Recombinant Protein A:
Contains four Fc-binding domains per protein
Better than Protein G for polyclonal IgG from rabbit, pig,
dog and cat serum
Poorer than Protein G for mouse IgG1, human IgG3 and
rat, goat and cow antibodies

Protein A.
Protein A is a cell wall component produced by

several strains ofStaphylococcus aureusthat

consists of a single polypeptide chain and
contains little or no carbohydrate. Recombinant
Protein A is produced inE. coliand functions
essentially the same as native Protein A. The
Protein A molecule contains four high-affinity
binding sites capable of interacting with the Fc
region from IgG of several species including
human and rabbit. Optimal binding occurs at pH
8.2, although binding is also effective at neutral
or physiological conditions (pH 7.0 to 7.6).

Protein A.
The interaction between Protein A and IgG

is not equivalent for all species. Even within

a species, Protein A interacts with some
subclasses of IgG and not others. For
instance, human IgG1, IgG2 and IgG4 bind
strongly, while IgG3 does not bind. There
are also many instances in which
monoclonal antibodies do not bind to
Protein A, especially the majority of rat
immunoglobulins and mouse IgG1.

Protein A.
Properties of Recombinant Protein A:

Source:E. coli
Molecular Weight: ~44,600 (Apparent MW
by SDS-PAGE: 45,000)
Form: salt-free powder
A280 of 0.1% solution: ~0.165
Isoelectric point (pI): 4.7-4.8
Related Products
Pierce Recombinant Protein A, Peroxidase
Conjugated

Protein A.
https://www.lifetechnologies.com/order/cat

alog/product/21184

Protein A.
https://tools.lifetechnologies.com/content

/sfs/manuals/MAN0011438_Pierce_Ig_Bind_Pro
teins_UG.pdf
https://tools.lifetechnologies.com/content/
sfs/brochures/TR0034-Ab-binding-proteins.pd
f

Protein A.
Crystal structure of aStaphylococcus

aureusprotein A domain complexed

with the Fab fragment of a human IgM
antibody: Structural basis for
recognition of B-cell receptors and
superantigen activity
Marc Graille et al. Proc Natl Acad Sci U S A.
2000 May 9; 97(10): 53995404.
PMCID:PMC25840
Immunology

Protein A.
Staphylococcus aureusproduces a

virulence factor, protein A (SpA), that

contains five homologous Ig-binding
domains. The interactions of SpA with the
Fab region of membrane-anchored Igs can
stimulate a large fraction of B cells,
contributing to lymphocyte clonal selection.

Protein A.
Staphylococcus aureusProtein A

Promotes Immune Suppression

Scott D. Kobayashi,
Frank R. DeLeo

Protein A.
SpA binds the Fc region of antibody and the Fab regions

of the B-cell receptor, processes that are known to

block opsonophagocytosis and cause B-cell deathin
vitro. In a recent study, Falugi et al. [F. Falugi, H. K. Kim,
D. M. Missiakas, and O. Schneewind, mBio 4(5):e0057513, 2013] showed that vaccination
withspamutantS.aureusstrains lacking antibody Fcand/or Fab-binding capacity protects against
subsequent challenge with the USA300 epidemic strain.
The findings provide strong support for the idea that
SpA promotesS.aureusimmune evasionin vivoand
form the foundation for a new approach in our efforts to
develop a vaccine that prevents
severeS.aureusinfections.

Autoimmune Diseases.
AUTOIMMUNE diseases, with the exception

of rheumatoid arthritis and autoimmune

thyroiditis, are individually rare, but
together they affect approximately 5
percent of the population in Western
countries.
Mechanisms of Autoimmunity:
Molecular Mimicry.
Superantigenic Stimulation.
Microbal Adjuvancticity.
Radiation

Autoimmune Diseases.
Endogenous:
Altered antigen presentation.
1. Loss of immunologic privilige.
2. Presentation of novel or crytic epitopes.
3. Alteration of self-antigen
4. Costumatory molecule expression
5. Cytokine production
B. Increased T cell help:
1. Cytokine production
2. Costumatory molecules

Autoimmune Diseases.
Increased B cell function
Apoptotic defects.
Cytokine imbalance.
Altered Immunoregulation.
Harrisons Principles of Internal Medicine.