54.

The extracellular matrix

Summary
The extracellular matrix is a network composed of mostly proteins and polysaccharides
in the space between the cells, but its water-based environment may, depending on tissue type,
also contain inorganic salts. It forms a scaffolding for the cells of tissues, and serves as a basis
for their movements, for example during embryonic development. The extracellular matrix is
in contact with the cytoskeleton through proteins of the cell membrane, and due to this also, it
has an influence on the shape of cells, and on signal transduction processes, which affect gene
regulation as well. Its main constituents are collagens, glycosaminoglycans and
proteoglycans, multiadhesive proteins all secreted by the cells, and integrins found in the
cell membrane.
Keywords: collagen, glycosaminoglycan, hyaluronan, proteoglycan, multiadhesive protein
integrin, focal adhesion, hemidesmosome, morphogenesis, signal transduction

The functions of the extracellular matrix

It primarily forms a scaffold to which cells can attach, providing a kind of foundation for
tissue structures, but its fibers also allow cells to move as they are attached to them. This
movement, for example, plays a major role during embryonic development, when stem cell
migration is crucial for the formation and shaping of tissues and organs, i.e. in morphogenesis.
The extracellular matrix is in physical contact with the cytoskeleton through cell membrane
proteins, thereby influencing cell shape. In tissues with a firmer matrix (e.g. bone, cartilage,
connective tissue of tendons and ligaments), cell shape and movement can be significantly
restricted. The association with cell membrane proteins, such as integrins, and through them
with the cytoskeleton, allows the manipulation of different intracellular signal transduction
pathways and thereby gene expression. But simply by the ability of extracellular matrix
elements to bind hormones and other ligands, they can modulate signal transduction processes
such as receptor binding and saturation.
The major components of the extracellular matrix are collagens, glycosaminoglycans and
proteoglycans, multiadhesive proteins released by secretion into the intercellular space, and
integrins transported into the cell membrane by a similar process (Figure 1).
 cadherin

cytoskeleton

integrin

cell membrane

multiadhesive matrix cell surface collagen fiber

protein proteoglycan proteoglycan

Figure 1. The main components of the extracellular matrix.

(Source: J. Szeberényi, M. Pap: Molecular cell biology Syllabus, 2024)

Collagens
Collagen proteins in humans are encoded by members of a large gene family and are the
most abundant proteins in the body overall. The individual polypeptide chains are extremely
flexible because every third amino acid is glycine, which is why they can unite in a triple-
helical (Figure 2) structure. The formation of hydrogen bonds between the three chains requires
the hydroxylation of the amino acids lysine and proline during maturation, for which the
presence of vitamin C is also necessary. In scurvy, the absence of vitamin C leads to the
breakdown of the resulting unstable triple helix so a general collagen deficiency occurs. The
consequences are the vulnerability of the skin, blood vessels, gums, gum bleeding, tooth loss
and slowed wound regeneration, etc.

Figure 2. The structure of collagen.

(left: a collagen polypeptide chain, Gly=glycine, X, Y: other amino acids. In the middle and
on the right: triple helical collagen structure).
(Source: www.priyamstudycentre.com, pdb101.rcsb.org/motm/4)
 In the rough endoplasmic reticulum, newly translated soluble procollagen chains are
glycosylated after successful hydroxylation, and the resulting triple helices are stabilized by
disulfide bridges. (Figure 3). Glycosylation is completed in the Golgi apparatus, and the N- and
C-terminal peptide regions of the protein that reaches the outside of the cell by exocytosis are
cleaved off. The resulting tropocollagen units are linked by covalent bonds to form collagen
fibers.

Figure 3. The steps of collagen synthesis.

(1: translation, the creation of soluble procollagen; 2: hydroxylation; 3: glycosylation; 4: the
formation of triple helical procollagen, the establishment of disulfide bridges; 5: vesicular
transport into the Golgi-apparatus, where glycosylation is finished; 6: exocytosis; 7: the
cleavage of the N- and C-terminal propeptides, the formation tropocollagen; 8: the linkage of
tropocollagen units through covalent bonds)
(Source: J. Szeberényi, M. Pap: Molecular cell biology Syllabus, 2024)
 These can then be arranged in parallel, forming strong bundles with high tensile strength in
the so-called fiber-forming collagens, which provide the physical resistance of bones, tendons
and ligaments. Mutations in their genes, often involving a glycine amino acid, lead to the
formation of unstable and consequently degradable triple helices. Mutations in bone collagens
are behind an inherited disease called osteogenesis imperfecta, which is associated with high
bone fragility. In Ehlers-Danlos syndrome, on the other hand, the mutation affects collagens of
the joints, skin and blood vessel walls, resulting in life-threatening arterial aneurysms
(vasodilatation) in addition to loose, stretchy joints and skin.
Collagen fibers are linked to elements of the extracellular matrix by fiber-associated
collagens. Network-forming collagens, together with elastic fibers composed of a protein
called elastin, form a thin layer of lamina basalis (basement membrane) to which epithelial cells
are attached.

Glycosaminoglycans and proteoglycan

Glycosaminoglycans are polysaccharides containing monotonically repeated
disaccharides, one of which is an N-acetyl glucosamine or an N-acetyl galactosamine and the
other is an acidic sugar, often with a sulfate group, for example. This special, highly hydrophilic
chemical composition gives them the ability to bind large amounts of water to form hydrated
gels, thus imparting a certain elasticity and consistency to various tissues, such as the connective
tissue of the skin or cartilage. The latter contains hyaluronan, which is present in a free form,
but often glycosaminoglycans such as dermatan sulfate, chondroitin sulfate and heparan sulfate
bind to proteins to compose proteoglycans. Of these, matrix proteoglycans are in the
intercellular space, such as the aggrecan of the cartilage tissue (Figure 4). Its central protein
chain (core protein) is associated with chondroitin sulfate and keratan sulfate molecules. These
units are then attached to a chain of hyaluronan through linking proteins to form large
(micrometer range) complexes with a brush-like appearance. Syndecan and fibroglycan, on the
other hand, are cell-surface proteoglycans, since their core proteins, which have a similar
structure to aggrecan, are transmembrane proteins at the same time (Figure 5).
 Figure 4. Aggrecan complex.
(Source: Lourdes Alcaide-Ruggiero et al.: Proteoglycans in Articular Cartilage and Their
Contribution to Chondral Injury and Repair Mechanisms, nt. J. Mol. Sci. 2023, 24(13),
10824)

syndecan

fibroglycan
heparan
sulfate

extracellular space chondroitin sulfate

cell membrane
cytosol
core protein

Figure 5. Cell surface proteoglycans.

(Source: J. Szeberényi, M. Pap: Molecular cell biology Syllabus, 2024)
Multiadhesive proteins
Multiadhesive proteins act as a linker between cell surface molecules, collagens,
glycosaminoglycans and proteoglycans. Examples include laminin, which stabilizes the
basement membrane of skin, capillaries, intestinal epithelium and kidney glomeruli, and
fibronectin, which enables connective tissue cells to bind to collagen.

Integrins
Integrins are transmembrane proteins with αβ heterodimeric structures that provide a
physical link between extracellular matrix elements and the cytoskeleton, ensuring the
integration of cells into tissues and their strong mechanical connection with their environment.
They form cell-matrix connections such as focal adhesions to microfilaments and
hemidesmosomes associated with intermediate filaments (Figure 6). In addition, integrins also
serve as mechanical receptors of the cell, capable of inducing intracellular signaling pathways
upon activation by various physical stimuli (pressure, traction, or even just the binding of
different extracellular matrix components to them).
fibroblast

epithelial cell

basal membrane

fibers of connective
tissue actin
filaments

cytokeratin
filaments
actin binding
multiprotein proteins
complex cytoplasmic plaque

cell membrane cell membrane

fibronectin integrin dimer laminin integrin dimer

fibers of connective
basal membrane
tissue

A. B.
Figure 6. The structure of a focal adhesion (A) and a hemidesmosome (B).
(Source: J. Szeberényi, M. Pap: Molecular cell biology Syllabus, 2024)
 Focal adhesions (Figure 6.A) are dynamic structures that allow different cells to attach to
elements of the extracellular matrix and to move by changing the location of these attachments.
The latter is made possible both by the cell's ability to easily disassemble focal adhesions and
then form a new one at a different point in the cell membrane, and by the fact that the
rearrangement of the actin network, which is also dynamic and bound to integrins via a looser
multiprotein complex, changes the shape of the cell. In this context, and in response to other,
for example, mechanical stimuli, integrins of focal adhesions can activate several signal
transduction pathways through the multiprotein complexes that bind to them, which will be
discussed in a later chapter.
Hemidesmosomes (Figure 6.B), on the other hand, are much more robust, strong and
stable, and help, for example, to anchor epithelial cells to the basement membrane, contributing
to the physical resistance of the upper layer of the skin. The denser structure of the protein
plaque intracellularly bound to the integrins of the hemidesmosomes and the attachment of
intermediate filaments that do not exhibit dynamic instability also support this static role.
The importance of integrins is demonstrated by experiments in mice in which the knockout
of certain integrin genes by KO mutation led to in utero (“in womb”) death of the animals.
Since, for example, during extravasation (see the chapter about cell-cell contacts), white blood
cells use integrins in addition to temporary, selectin-based cell-cell contacts to bind more stably
to the appropriate site on the inner surface of the capillary, it is understandable that the
syndrome called leukocyte adhesion deficiency associated with recurrent infections may be
caused not only by mutations in selectin but also in certain integrin genes.
For most cells, attachment to the extracellular matrix is essential for their survival, mainly
because of the importance of intracellular signal transduction pathways initiated by activated
integrins that inhibit cell death (see later chapter on integrin signaling). However, in some tumor
cells, altered integrin expression, such as mutant or abnormal αβ-complex integrins, can
stimulate these signaling pathways important for cell survival even when the cell is detached
from its tissue environment. This allows the tumor to invade surrounding tissues or to
metastasize into a distant organ.
Questions

Name the most important structural components of the extracellular matrix (5)!
collagen fibers, glycosaminoglycans, proteoglycans, multiadhesive proteins, integrins

Give three examples for the function of the extracellular matrix!

attachment site for cells, involvement in signal transduction processes, influencing cell shape,
structural base of tissues, etc.

What is the molecular background of scurvy caused by decreased vitamin C uptake?

hydroxylation of collagen is decreased

What is the name of transmembrane proteins acting as receptors for the extracellular matrix?
integrins