Linear epitope
Encyclopedia
A linear or a sequential epitope is an epitope
that is recognized by antibodies
by its linear sequence of amino acid
s, or primary structure
. In contrast, most antibodies recognize a conformational epitope
that has a specific three-dimensional shape and its protein structure
.
An antigen is any substance that the immune system
can recognize as foreign. Since, antigens are usually proteins that are too large to bind as a whole to any receptor, only specific segments that form the antigen bind with a specific antibody. Such segments are called epitope
s. Likewise, it is only paratope
of the antibody that comes in contact with the epitope.
Protein
s are composed of repeating nitrogen
-containing subunits called amino acid
s. The linear sequence of amino acids that compose a protein is called its primary structure
, which does not exist in nature. But, when an antigen is broken down in a lysosome
, it yields small peptide
s, which can be recognized through the amino acids that lie continuously in a line, and hence are called linear epitopes.
, immunohistochemistry
, and ELISA
, one should carefully choose antibodies that recognize linear or conformational epitopes.
For instance, if a protein
sample is boiled, treated with beta-mercaptoethanol, and run in SDS-PAGE
for the Western blot, the proteins are essentially denatured
and therefore cannot assume their natural three-dimensional conformations. Therefore, antibodies that recognize linear epitopes instead of conformational epitopes are chosen for immunodetection. In contrast, in immunohistochemistry where protein structure is preserved, antibodies are preferred that recognize conformational epitopes.
Epitope
An epitope, also known as antigenic determinant, is the part of an antigen that is recognized by the immune system, specifically by antibodies, B cells, or T cells. The part of an antibody that recognizes the epitope is called a paratope...
that is recognized by antibodies
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...
by its linear sequence of amino acid
Amino acid
Amino acids are molecules containing an amine group, a carboxylic acid group and a side-chain that varies between different amino acids. The key elements of an amino acid are carbon, hydrogen, oxygen, and nitrogen...
s, or primary structure
Primary structure
The primary structure of peptides and proteins refers to the linear sequence of its amino acid structural units. The term "primary structure" was first coined by Linderstrøm-Lang in 1951...
. In contrast, most antibodies recognize a conformational epitope
Conformational epitope
A conformational epitope is a sequence of subunits composing an antigen that come in direct contact with a receptor of the immune system.An antigen is any substance that the immune system can recognize as foreign...
that has a specific three-dimensional shape and its protein structure
Protein structure
Proteins are an important class of biological macromolecules present in all organisms. Proteins are polymers of amino acids. Classified by their physical size, proteins are nanoparticles . Each protein polymer – also known as a polypeptide – consists of a sequence formed from 20 possible L-α-amino...
.
An antigen is any substance that the immune system
Immune system
An immune system is a system of biological structures and processes within an organism that protects against disease by identifying and killing pathogens and tumor cells. It detects a wide variety of agents, from viruses to parasitic worms, and needs to distinguish them from the organism's own...
can recognize as foreign. Since, antigens are usually proteins that are too large to bind as a whole to any receptor, only specific segments that form the antigen bind with a specific antibody. Such segments are called epitope
Epitope
An epitope, also known as antigenic determinant, is the part of an antigen that is recognized by the immune system, specifically by antibodies, B cells, or T cells. The part of an antibody that recognizes the epitope is called a paratope...
s. Likewise, it is only paratope
Paratope
The paratope is the part of an antibody which recognises an antigen, the antigen-binding site of an antibody. It is a small region of the antibody's Fv region and contains parts of the antibody's heavy and light chains....
of the antibody that comes in contact with the epitope.
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...
s are composed of repeating nitrogen
Nitrogen
Nitrogen is a chemical element that has the symbol N, atomic number of 7 and atomic mass 14.00674 u. Elemental nitrogen is a colorless, odorless, tasteless, and mostly inert diatomic gas at standard conditions, constituting 78.08% by volume of Earth's atmosphere...
-containing subunits called amino acid
Amino acid
Amino acids are molecules containing an amine group, a carboxylic acid group and a side-chain that varies between different amino acids. The key elements of an amino acid are carbon, hydrogen, oxygen, and nitrogen...
s. The linear sequence of amino acids that compose a protein is called its primary structure
Primary structure
The primary structure of peptides and proteins refers to the linear sequence of its amino acid structural units. The term "primary structure" was first coined by Linderstrøm-Lang in 1951...
, which does not exist in nature. But, when an antigen is broken down in a lysosome
Lysosome
thumb|350px|Schematic of typical animal cell, showing subcellular components. [[Organelle]]s: [[nucleoli]] [[cell nucleus|nucleus]] [[ribosomes]] [[vesicle |vesicle]] rough [[endoplasmic reticulum]]...
, it yields small peptide
Peptide
Peptides are short polymers of amino acid monomers linked by peptide bonds. They are distinguished from proteins on the basis of size, typically containing less than 50 monomer units. The shortest peptides are dipeptides, consisting of two amino acids joined by a single peptide bond...
s, which can be recognized through the amino acids that lie continuously in a line, and hence are called linear epitopes.
Significance
While performing molecular assays involving use of antibodies such as in the Western blotWestern blot
The western blot is a widely used analytical technique used to detect specific proteins in the given sample of tissue homogenate or extract. It uses gel electrophoresis to separate native proteins by 3-D structure or denatured proteins by the length of the polypeptide...
, immunohistochemistry
Immunohistochemistry
Immunohistochemistry or IHC refers to the process of detecting antigens in cells of a tissue section by exploiting the principle of antibodies binding specifically to antigens in biological tissues. IHC takes its name from the roots "immuno," in reference to antibodies used in the procedure, and...
, and ELISA
ELISA
Enzyme-linked immunosorbent assay , is a popular format of a "wet-lab" type analytic biochemistry assay that uses one sub-type of heterogeneous, solid-phase enzyme immunoassay to detect the presence of a substance in a liquid sample."Wet lab" analytic biochemistry assays involves detection of an...
, one should carefully choose antibodies that recognize linear or conformational epitopes.
For instance, if a 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...
sample is boiled, treated with beta-mercaptoethanol, and run in SDS-PAGE
SDS-PAGE
SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis, describes a collection of related techniques widely used in biochemistry, forensics, genetics and molecular biology to separate proteins according to their electrophoretic mobility...
for the Western blot, the proteins are essentially denatured
Denaturation (biochemistry)
Denaturation is a process in which proteins or nucleic acids lose their tertiary structure and secondary structure by application of some external stress or compound, such as a strong acid or base, a concentrated inorganic salt, an organic solvent , or heat...
and therefore cannot assume their natural three-dimensional conformations. Therefore, antibodies that recognize linear epitopes instead of conformational epitopes are chosen for immunodetection. In contrast, in immunohistochemistry where protein structure is preserved, antibodies are preferred that recognize conformational epitopes.