Protein A
Encyclopedia
Protein A is a 40-60 kDa
MSCRAMM
surface protein
originally found in the cell wall of the bacterium Staphylococcus aureus
. It is encoded by the spa gene and its regulation is controlled by DNA topology, cellular osmolarity, and a two-component system called ArlS-ArlR. It has found use in biochemical research because of its ability to bind immunoglobulins. 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 of mammalian species, most notably IgGs. It binds the heavy chain with the Fc region of most immunoglobulins and also within the Fab region in 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 antibody
function), the bacteria disrupts opsonization and phagocytosis.
Protein A binds with moderate affinity to human IgM
, IgA
and IgE
as well as to mouse IgG3 and IgG1.
It does not react with human IgG3 or IgD
, nor will it react to mouse IgM, IgA or IgE.
The capacity of protein A to bind antibodies with such high affinity is the driving motivation for its industrial scale use in biologic pharmaceuticals. The protein A used for production of antibodies in bio-pharmaceuticals is most commonly bound to a stationary phase chromatography resin.
, Protein A/G
and Protein L
are all commonly used to purify, immobilize or detect immunoglobulins.
Protein A is often coupled to other molecules such as a fluorescent dye, enzyme
s, biotin
, colloidal gold
or radioactive iodine without affecting the antibody binding site. It is also widely utilized coupled to magnetic, latex and agarose beads.
Protein A is often immobilized onto a solid support and used as reliable method for purifying total IgG from crude protein mixtures such as serum
or ascites
fluid, or coupled with one of the above markers to detect the presence of antibodies. Immunoprecipitation
studies with protein A conjugated to beads are also commonly used to purify proteins or protein complexes indirectly through antibodies against the protein or protein complex of interest.
Atomic mass unit
The unified atomic mass unit or dalton is a unit that is used for indicating mass on an atomic or molecular scale. It is defined as one twelfth of the rest mass of an unbound neutral atom of carbon-12 in its nuclear and electronic ground state, and has a value of...
MSCRAMM
MSCRAMM
MSCRAMM adhesin proteins mediate the initial attachment of bacteria to host tissue, providing a critical step to establish infection....
surface protein
Protein
Proteins are biochemical compounds consisting of one or more polypeptides typically folded into a globular or fibrous form, facilitating a biological function. A polypeptide is a single linear polymer chain of amino acids bonded together by peptide bonds between the carboxyl and amino groups of...
originally found in the cell wall of the bacterium Staphylococcus aureus
Staphylococcus aureus
Staphylococcus aureus is a facultative anaerobic Gram-positive coccal bacterium. It is frequently found as part of the normal skin flora on the skin and nasal passages. It is estimated that 20% of the human population are long-term carriers of S. aureus. S. aureus is the most common species of...
. It is encoded by the spa gene and its regulation is controlled by DNA topology, cellular osmolarity, and a two-component system called ArlS-ArlR. It has found use in biochemical research because of its ability to bind immunoglobulins. 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 of mammalian species, most notably IgGs. It binds the heavy chain with the Fc region of most immunoglobulins and also within the Fab region in 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 antibody
Antibody
An antibody, also known as an immunoglobulin, is a large Y-shaped protein used by the immune system to identify and neutralize foreign objects such as bacteria and viruses. The antibody recognizes a unique part of the foreign target, termed an antigen...
function), the bacteria disrupts opsonization and phagocytosis.
Protein A antibody binding
Protein A binds with high affinity to human IgG1 and IgG2 as well as mouse IgG2a and IgG2b.Protein A binds with moderate affinity to human IgM
IGM
IGM as an acronym or abbreviation can refer to:* Immunoglobulin M , the primary antibody against A and B antigens on red blood cells* International Grandmaster, a chess ranking* intergalactic medium* Intragroup medium - see: Intracluster medium...
, IgA
IGA
Iga or IGA may stand for:-Given name:* a female given name of Polish origin. The name originates from the female given name Jadwiga and stands for gia,or gina in the USA....
and IgE
IGE
IGE was one of the largest services company buying and selling virtual currencies and accounts for MMORPG. During its peak time, it had offices in Los Angeles, China , and headquarters & customer service centre in Hong Kong. IGE was one of the main monopoly in virtual economy services, also known...
as well as to mouse IgG3 and IgG1.
It does not react with human IgG3 or IgD
IGD
IGD can refer to:*Internet Gateway Device as defined in UPnP.*İGD, İlerici Gençler Derneği, Progressive Young Association of Turkey*Immunoglobulin D, an antibody protein involved in the maturation of B cells....
, nor will it react to mouse IgM, IgA or IgE.
The capacity of protein A to bind antibodies with such high affinity is the driving motivation for its industrial scale use in biologic pharmaceuticals. The protein A used for production of antibodies in bio-pharmaceuticals is most commonly bound to a stationary phase chromatography resin.
Other antibody binding proteins
In addition to Protein A, other immunoglobulin-binding bacterial proteins such as Protein GProtein G
Protein G is an immunoglobulin-binding protein expressed in group C and G Streptococcal bacteria much like Protein A but with differing specificities. It is a 65-kDa and a 58 kDa cell surface protein that has found application in purifying antibodies through its binding to the Fc region...
, Protein A/G
Protein A/G
Protein A/G is a recombinant fusion protein that combines IgG binding domains of both Protein A and Protein G. Protein A/G contains four Fc binding domains from Protein A and two from Protein G, yielding a final mass of 50,460 daltons...
and Protein L
Protein L
Protein L was first isolated from the surface of bacterial species Peptostreptococcus magnus and was found to bind immunoglobulins through L chain interaction, from which the name was suggested. It consists of 719 amino acid residues...
are all commonly used to purify, immobilize or detect immunoglobulins.
Role in pathogenesis
As a pathogen Staphylococcus aureus utilizes Protein A, along with a host of other proteins and surface factors to aid its survival and, thus, virulence. Protein A helps inhibit phagocytic engulfment and acts as an immunological disguise. Mutants of S. aureus lacking protein A are more efficiently phagocytosed in vitro, and mutants in infection models have diminished virulence.Research
Recombinant Staphylococcal Protein A is often produced in E. coli for use in immunology and other biological research. One recombinant form of Protein A is called MabSelect.Protein A is often coupled to other molecules such as a fluorescent dye, enzyme
Enzyme
Enzymes are proteins that catalyze chemical reactions. In enzymatic reactions, the molecules at the beginning of the process, called substrates, are converted into different molecules, called products. Almost all chemical reactions in a biological cell need enzymes in order to occur at rates...
s, biotin
Biotin
Biotin, also known as Vitamin H or Coenzyme R, is a water-soluble B-complex vitamin discovered by Bateman in 1916. It is composed of a ureido ring fused with a tetrahydrothiophene ring. A valeric acid substituent is attached to one of the carbon atoms of the tetrahydrothiophene ring...
, colloidal gold
Colloidal gold
Colloidal gold is a suspension of sub-micrometre-sized particles of gold in a fluid — usually water. The liquid is usually either an intense red colour , or a dirty yellowish colour ....
or radioactive iodine without affecting the antibody binding site. It is also widely utilized coupled to magnetic, latex and agarose beads.
Protein A is often immobilized onto a solid support and used as reliable method for purifying total IgG from crude protein mixtures such as serum
Blood serum
In blood, the serum is the component that is neither a blood cell nor a clotting factor; it is the blood plasma with the fibrinogens removed...
or ascites
Ascites
Ascites is a gastroenterological term for an accumulation of fluid in the peritoneal cavity.The medical condition is also known as peritoneal cavity fluid, peritoneal fluid excess, hydroperitoneum or more archaically as abdominal dropsy. Although most commonly due to cirrhosis and severe liver...
fluid, or coupled with one of the above markers to detect the presence of antibodies. Immunoprecipitation
Immunoprecipitation
Immunoprecipitation is the technique of precipitating a protein antigen out of solution using an antibody that specifically binds to that particular protein. This process can be used to isolate and concentrate a particular protein from a sample containing many thousands of different proteins...
studies with protein A conjugated to beads are also commonly used to purify proteins or protein complexes indirectly through antibodies against the protein or protein complex of interest.