Collagen

Collagen is the main structural protein in the extracellular matrix of the connective tissues of many animals. It is the most abundant protein in mammals, making up 25% to 35% of protein content. Amino acids are bound together to form a triple helix of elongated fibril known as a collagen helix. It is mostly found in cartilage, bones, tendons, ligaments, and skin. Vitamin C is vital for collagen synthesis.
Depending on the degree of mineralization, collagen tissues may be rigid or compliant or have a gradient from rigid to compliant. Collagen is also abundant in corneas, blood vessels, the gut, intervertebral discs, and dentin. In muscle tissue, it serves as a major component of the endomysium. Collagen constitutes 1% to 2% of muscle tissue and 6% by weight of skeletal muscle. The fibroblast is the most common cell creating collagen in animals. Gelatin, which is used in food and industry, is collagen that was irreversibly hydrolyzed using heat, basic solutions, or weak acids.

Etymology

The word collagen comes from Greek κόλλα, kólla 'glue' and the suffix -γέν, -gen 'producing'.

Types

As of 2011, 28 types of human collagen have been identified, described, and classified according to their structure. This diversity shows collagen's diverse functionality. All of the types contain at least one triple helix. Over 90% of the collagen in humans is Type I and Type III collagen.

Fibrillar
Non-fibrillar
* FACIT
* Short-chain
* Basement membrane
* Multiplexin
* MACIT
* Microfibril-forming
* Anchoring fibrils

The five most common types are:

Type I: skin, tendon, vasculature, organs, bone
Type II: cartilage
Type III: reticulate, commonly found alongside type I
Type IV: forms basal lamina, the epithelium-secreted layer of the basement membrane
Type V: cell surfaces, hair, and placenta
In humans

Cardiac

The collagenous cardiac skeleton, which includes the four heart valve rings, is histologically, elastically and uniquely bound to cardiac muscle. The cardiac skeleton also includes the separating septa of the heart chambers – the interventricular septum and the atrioventricular septum. Collagen contribution to the measure of cardiac performance summarily represents a continuous torsional force opposed to the fluid mechanics of blood pressure emitted from the heart. The collagenous structure that divides the upper chambers of the heart from the lower chambers is an impermeable membrane that excludes both blood and electrical impulses through typical physiological means. With support from collagen, atrial fibrillation never deteriorates to ventricular fibrillation. Collagen is layered in variable densities with smooth muscle mass. The mass, distribution, age, and density of collagen all contribute to the compliance required to move blood back and forth. Individual cardiac valvular leaflets are folded into shape by specialized collagen under variable pressure. Gradual calcium deposition within collagen occurs as a natural function of aging. Calcified points within collagen matrices show contrast in a moving display of blood and muscle, enabling methods of cardiac imaging technology to arrive at ratios essentially stating blood in and blood out. Pathology of the collagen underpinning of the heart is understood within the category of connective tissue disease.

Bone grafts

As the skeleton forms the structure of the body, it is vital that it maintains its strength and its structure, even after breaks and injuries. Collagen is used in bone grafting because its triple-helix structure makes it a very strong molecule. It is ideal for use in bones, as it does not compromise the structural integrity of the skeleton. The triple helical structure prevents collagen from being broken down by enzymes, it enables adhesiveness of cells, and it is important for the proper assembly of the extracellular matrix.

Tissue regeneration

Collagen scaffolds are used in tissue regeneration, whether in sponges, thin sheets, gels, or fibers. Collagen has favorable properties for tissue regeneration, such as pore structure, permeability, hydrophilicity, and stability in vivo. Collagen scaffolds also support deposition of cells, such as osteoblasts and fibroblasts, and once inserted, facilitate growth to proceed normally.

Reconstructive surgery

Collagens are widely used in the construction of artificial skin substitutes used for managing severe burns and wounds. These collagens may be derived from cow, horse, pig, or even human sources; and are sometimes used in combination with silicones, glycosaminoglycans, fibroblasts, growth factors, and other substances.

Wound healing

Collagen is one of the body's key natural resources and a component of skin tissue that can benefit all stages of wound healing. When collagen is made available to the wound bed, closure can occur. This avoids wound deterioration and procedures such as amputation.
Collagen is used as a natural wound dressing because it has properties that artificial wound dressings do not have. It resists bacteria, which is vitally important in wound dressing. As a burn dressing, collagen helps it heal fast by helping granulation tissue to grow over the burn.
Throughout the four phases of wound healing, collagen performs the following functions:

Guiding: collagen fibers guide fibroblasts because they migrate along a connective tissue matrix.
Chemotaxis: collagen fibers have a large surface area which attracts fibrogenic cells which help healing.
Nucleation: in the presence of certain neutral salt molecules, collagen can act as a nucleating agent causing formation of fibrillar structures.
Hemostasis: Blood platelets interact with the collagen to make a hemostatic plug.
Use in basic research

Collagen is used in laboratory studies for cell culture, studying cell behavior and cellular interactions with the extracellular environment. Collagen is also widely used as a bioink for 3D bioprinting and biofabrication of 3D tissue models.

Biology

The collagen protein is composed of a triple helix, which generally consists of two identical chains and an additional chain that differs slightly in its chemical composition. The amino acid composition of collagen is atypical for proteins, particularly with respect to its high hydroxyproline content. The most common motifs in collagen's amino acid sequence are glycine-proline-X and glycine-X-hydroxyproline, where X is any amino acid other than glycine, proline or hydroxyproline.
The table below lists average amino acid composition for fish and mammal skin.

Amino acid	Abundance in mammal skin	Abundance in fish skin
Glycine	329	339
Proline	126	108
Alanine	109	114
Hydroxyproline	95	67
Glutamic acid	74	76
Arginine	49	52
Aspartic acid	47	47
Serine	36	46
Lysine	29	26
Leucine	24	23
Valine	22	21
Threonine	19	26
Phenylalanine	13	14
Isoleucine	11	11
Hydroxylysine	6	8
Methionine	6	13
Histidine	5	7
Tyrosine	3	3
Cysteine	1	1
Tryptophan	0	0

Synthesis

First, a three-dimensional stranded structure is assembled, mostly composed of the amino acids glycine and proline. This is the collagen precursor procollagen. Then, procollagen is modified by the addition of hydroxyl groups to the amino acids proline and lysine. This step is important for later glycosylation and the formation of collagen's triple helix structure. Because the hydroxylase enzymes performing these reactions require vitamin C as a cofactor, a long-term deficiency in this vitamin results in impaired collagen synthesis and scurvy. These hydroxylation reactions are catalyzed by the enzymes prolyl 4-hydroxylase and lysyl hydroxylase. The reaction consumes one ascorbate molecule per hydroxylation. Collagen synthesis occurs inside and outside cells.
The most common form of collagen is fibrillary collagen. Another common form is meshwork collagen, which is often involved in the formation of filtration systems. All types of collagen are triple helices, but differ in the make-up of their alpha peptides created in step 2. Below we discuss the formation of fibrillary collagen.

Transcription of mRNA: Synthesis begins with turning on genes associated with the formation of a particular alpha peptide. About 44 genes are associated with collagen formation, each coding for a specific mRNA sequence, and are typically named with the "COL" prefix.
Pre-pro-peptide formation: The created mRNA exits the cell nucleus into the cytoplasm. There, it links with the ribosomal subunits and is translated into a peptide. The peptide goes into the endoplasmic reticulum for post-translational processing. It is directed there by a signal recognition particle on the endoplasmic reticulum, which recognizes the peptide's signal sequence. The processed product is a pre-pro-peptide called preprocollagen.
Pro-collagen 'formation: Three modifications of the pre-pro-peptide form the alpha peptide:
# The signal peptide on the N-terminal is removed. This molecule is called 'propeptide.'
# Lysines and prolines are hydroxylated by the enzymes 'prolyl hydroxylase' and 'lysyl hydroxylase', producing hydroxyproline and hydroxylysine. This helps in cross-linking the alpha peptides. This enzymatic step requires vitamin C as a cofactor. In scurvy, the lack of hydroxylation of prolines and lysines causes a looser triple helix.
# Glycosylation occurs by adding either glucose or galactose monomers onto the hydroxyl groups that were placed onto lysines, but not on prolines.
# Three of the hydroxylated and glycosylated propeptides twist into a triple helix, forming procollagen. It is packaged into a transfer vesicle destined for the Golgi apparatus.
Modification and secretion: In the Golgi apparatus, the procollagen goes through one last post-translational modification, adding oligosaccharides. Then it is packaged into a secretory vesicle to be secreted from the cell.
Tropocollagen formation: Outside the cell, membrane-bound enzymes called collagen peptidases remove the unwound ends of the molecule, producing tropocollagen. Defects in this step produce various collagenopathies called Ehlers–Danlos syndrome. This step is absent when synthesizing type III, a type of fibrillar collagen.
Collagen fibril formation': Lysyl oxidase, a copper-dependent enzyme, acts on lysines and hydroxylysines, producing aldehyde groups, which eventually form covalent bonds between tropocollagen molecules. This polymer of tropocollagen is called a collagen fibril.