Biochemistry of skin

Jana Novotn
Department of Med. Biochemistry
2nd Faculty of Medicine
Charles University

Skin

it provides barrier against a range of noxious stress (UV irradiation,

mechanical, chemical and biological insults);
acts as the periphery sensing system;
system which maintaining body homeostasis.
2 m2 in area
~ 2.5 mm thick on average
constitutes 6% of our total body weight (5 6 kg)
barrier to prevent a desiccation and temperature balance
protection to the UV radiation absorbing pigmentation system
complex immuno-regulatory network protection
normal skin pH is somewhat acidic - the range of 4.2. to 5.6.

Human skin layers

Mammalian skin is composed of two primary layers:

the epidermis, which provides waterproofing and serves as a
barrier to infection;
the dermis is responsible for the tensile strength of skin. Its main
functions are to regulate temperature and to supply the
epidermis with nutrient-saturated blood. Much of the body's
water supply is stored within the dermis.

Epidermis
An external stratified, non-vasularized
epithelium (75 150 m thick),
continually keratinizing
Stratum corneum 15 30 sheet
of non-viable, but biochemicaly
active corneocytes
Stratum granulosum 3 5 sheet
of non-dividing keratinocytes,
producing keratino-hyalin
Stratum spinosum 8 10 sheet of
keratinocytes with limited dividing
capacity, Langerhans cells
Stratum basale maturing/aging
keratinocytes, melanocytes, Merkel
cells (receptor cells)

Keratins
keratinocytes contain filaments of the keratin
intermediate filament (KIF) family (cytoskeleton)
hair, nails, horny layers of the skin are
formed from keratin cytoskeleton of dead
cells.
two primary groups of keratins, the keratins and the -keratins
-keratins occur in mammals, -keratins in
birds, reptiles
both form are right handed helical structure
2 types
type I acidic keratins
type II basic keratins
heterodimer type I forming a coild coil with type II

Composition and Structure of

Keratin

Human skin contains ~ 20 genetically

different keratins
Long stretches -helix is interrupted by
short non-helical segments
The most abundant amino acid are
glycine and alanine, cysteine can account
for up to 24%
Contact between 2 helices are formed
by hydrophobic amino acid side chain on
1 edge of each helix
two polypeptides form a dimeric colid coil
protofilaments are formed from two
staggered rows of head-to-tail associated
coils
protofilaments dimerize to form a protofibril
and four of which form a microfibril

http://biochem118.stanford.edu/Papers/Protein%20Papers/Voet%26Voet%20chapter6.pdf

Composition and Structure of

Keratin
Intra- and intermolecular hydrogen bonds, disufid bridges
occure at all keratins.
In cells, keratin type I forms pair with keratine type II
Different keratin types are expressed in different cell types
and different layers of epidermis:
cytoskeleton of epithelial cells - K14 (type I) & K5 (type
II), K18 (type I) & K8 (type II)
Basal layer K13 (type I) & K4 (type II)
Spinus and granular layer K10 (type I) & K1 (type II)
Stratum corneum K3 (type I) & K12 (type II)
Hairs and nails various other keratin pairs

The Epidermal Permeability

Barrier
Barrier function in human epidermis depends on
transglutaminase-mediated cross-linking of structural
proteins and lipids (biological glues)
post-translation modification of proteins formation of covalent bond
between a -amino group of lysine and the free amine group of
glutamine.

Bonds formed by transglutaminase exhibit high resistance to

proteolytic degradation.
Proteins are than highly resistent to mechanical perturbation
and proteolysis.
The quality of the S. corneum barrier depends on the
presence of equimolar concentration of ceramides,
cholesterol and fatty acids.
Changes in the concentration of any of these can affect
barrier quality.

Fatty Acids in Epidermis

Arachidonic acid and 20-carbon PUFA can be
metabolized by either cyclooxygenase or lipoxygenase
pathways prostaglandins, hydroxyeicosatetraenoic
acids.
phospholipids are starting point for the arachidonic
acid pathway during inflammation (allergic reaction)
Some of these metabolites can interact with signaling
system in proliferating and differentiating epidermal cells
modulation of protein kinase C, nuclear MAP-kinase

Epidermal Cell Differentiation

and Turnover
Basal keratinocytes transformation ~ 30 days to
corneocytes.
Damage cells are removed by normal squamation.
Genetic damage - (UV-R) trigger apoptosis (within
hours) sunburn cells.
Skin protection against UV-R concentrating transferred
melanin over vulnerable keratocyte nucleus.
Other insults can induce keratnocyte apoptosis
chemical, mechanical, immunological.
The principal marker for keratinocyte/epidermal
differentiation is expression of particular keratin pairs.

Epidermal Cell Differentiation

and Turnover
Majority of over 30 keratins currently known.
Proliferative basal keratinocytes express K5 and K14;
keratinocytes in the early stages of maturation/differentiation
switch to K1 and K10.
The pluri-potent stem cells for keratinocytes sebaceous
gland and epidermis rised from hair folicules.
Ca2+ plays pivotal role in epidermal differentiation - 4-fold
increase of extracellular Ca2+ in S. corneum.
Keratinocyte differentiation is regulated by hormones and
vitamins - D3 and retinol from diet, thyroid hormones and
steroid hormones.
The skin has nucleas receptors for glucocorticoids, estrogen,
androgen and progesterone.

Epidermal Cell Differentiation

and Turnover

Importan factors for keratocyte differentiation are Ca2+-dependent

integrins the receptors for the extracellular matrix fibronectin
binding.
Laminin and collagen IV and VII (basemen membrane components)
regulation of keratinocytes migration on basement membrane
(very important during wound healing).
Migrating keratinocytes produce many matrix metalloproteinases.
Mature keratinocytes (in S. graulosum) contain protein-rich,
keratohyalin granules and lipid-rich, lamellar granules.
Lipids from lamelar granules form the sheets of the lipid permeability
of the lipid permeability barrier of the epidermis.

Melanocytes

Melanocytes are melanin-producing cells located in S. basale

Precursor - melanoblast
Melanin is stored in the melanosomes.
Epidermal melanin unit - the anatomical relationship between
keratinocytes and melanocytes.
1 melanocyte is in contact with ~ 40 keratinocytes
Melanocytes extend arms to transfer melanosomes into the
keratinocytes

Melanocytes
Cover picture:The Rab27a
GTPase associates with
melanosomes and regulates
their transport to, and retention
in, the peripheral cytoplasm in
skin melanocytes.
Melanosome transport also
requires the microtubule and
actin cytoskeleton. Staining of
a cultured murine melanocyte
for filamentous actin (red) and
microtubules (blue) reveals a
close relationship between
Rab27a-labelled melanosomes
(green) and these cytoskeletal
elements.
Hume et al. JCB 2001;152 (4): 795

Formation of melanosomes

Melanosomes - elliptic membranebound organelles (melanin

synthesis).
Synthesis of matrix proteins and
tyrosinase (TYR) on the rough
endoplasmic reticulum.
TYR undergoes post translational
modification in the form of
glycosylation in the Golgi apparatus.
Fusion of premelanosomes with
coated vesicles containing tyrosinase
- formation of the melanosome.
Melanosome migrates into one of the
dendrites of the melanocyte
transfer to a neighboring
keratinocyte.

Production of Melanin
Three enzymes in melanosomes whih absolutely required for
different melanin type synthesis
tyrosinase (TYR) responsible for critical step of melanogenesis (tyrosine
hydroxylation)
tyrosinase-related protein 1 (TYR1) and DOPAchrome tautomerase

(DHI = 5,6-dihydroxyindole; DHICA = 5,6-dihydroxyindole-2carboxylic acid)

Melanins

Melanins are polymorphous and multifunctional polymers of

eumelanin, pheomelanin, mixed melanins (a combination of the
two); and neuromelanin
Mammalian cells produce black-brown eumelanin and yellowredish pheomelanin
Eumelanin - highly heterogenous polymer consisting of DHI and
DHICA units in reduced or oxidized states.
Pheomelanin - mainly sulfur-containing benzothiazine derivatives.
Neuromelanin is produces in dopaminergic neurons of substantia
nigra.
Melanin absorbs UV light at a wavelength of 280 - 320 nm
Both eumelanin and pheomelanin play important protective role in
binding to cations, anions, drugs, chemicals, etc.

Factors Involve in Melanin

Production

The melanin granules accumulate above the nuclei of keratinocytes

and absorb harmful UV-R before it can reach the nucleus and
damage the DNA.

Quick responds of the melanocyte-keratinocyte complex to a wide

range of environmental stimuli (paracrine and/or autocrine) - to UVR, melanocyte-stimulating hormone (MSH), endothelins, growth
factors, cytokines, etc.

UV-R exposure melanocytes increase their expression of

proopiomelanocortin (POMC, the precursor of MSH) and its receptor
melanocortin 1 receptor (MC1-R), TYR and TYRP1, protein kinase
C (PKC), and other signaling factors.

Factors Involve in Melanin

Production
Fibroblasts (possibly other cells in skin) - produce
cytokines, growth factors, and inflammatory mediators that
can increase melanin production and/or stimulate melanin
transfer to keratinocytes by melanocytes.
Other factors derived from keratinocytes which can
regulate proliferation and/or differentiation of melanocytes:
-MSH, ACTH, basic fibroblast growth factor (FGF),
nerve growth factor (NGF), endothelins, granulocytemacrophage colony-stimulating factor (GM-CSF),
leukemia inhibitory factor (LIF), and hepatocyte growth
factor (HGF).

Other Epidermal Cells

Langerhans cells - dendritic cells - arise from bone marrow early in

embryonic development, occupie 2 - 8% of epidermis
important element of the immune system, interacting with T-cells
resided in suprabasal layer - attracted to keranocytes by Ecadherin receptor
their motion is regulated by specific integrin receptor and by
TNF
in the stratum germinativum interacts with the allergen and migrates
to the lymphoid gland - then teache the T cells about the allergen
interact specifically with T-lymphocytes and keratinocytes to initiate
host response to antigens (allergens)
UV B stimulates synthesis and release of TNF- by keratinocytes
which in turn modifies the behavior and morphology of Langerhans
cells, decreases their total number.

Langehans
cell

Allergen
cytokine

T cell

Activated
T cell

Other Epidermal Cells

Merkel cells location in S. germinativum
have synaptic contacts with sensory nerve endings
associated with the sense of light touch discrimination
of shapes and textures.

Dermis

responsible for the tensile strength of skin

main functions regulation of temperature and to supply the
epidermis with nutrient
much of the body's water supply is stored within the dermis
components:

connective tissue
hair follicles
sweat glands
sebaceous or oil glands
apocrine glands
lymph vessels
blood vessels

The main cell type - fibroblast

Dermal Proteins and Extracellular

Matrix
Collagen about 90% of total dermal proteins
predominatly type I (85 90%),
type III (8 -11%),
minor type V (2 4%), (papillary dermis, matrix around vessels,
nerves),
type VI associated with fibrils and interfibrillar spaces
(responsible for fine structure in early prenatal development of
skin).

Elastin, proteoglycans, glycoproteins, water and

hyaluronic acid
Collagen structure - refer to lecture on Collagens
Elastin, proteoglycans, glycoproteins refer to lecture on
Extracellular matrix

Skin Appendages
Skin plays in the body homeostasis, therefore is
well-equiped with secretory (release of chemicals
from cells for physiological function) and excretory
(elimination of weste products of metabolism) capacity.
sweat glands [can be sweat secreted with strong
odour (apocrine) or with a faint odour (eccrine)].
sebaceous glands (secrete sebum onto hair follicle to
oil the hair).
hair follicle

Sweat Glands
3 4 million eccrine sweat glands are
in our skin each producing water
perspiration (serves mainly to cool us)
and maintain core temperature at
37.5oC.
At maximum output the eccrine sweat
glands can excrete as much as 3 l/hour,
and heat loss is more than 18 kcal min1.
Humans utilize eccrine sweat glands as
primary form of cooling.
Apocrine sweat glands are larger, have
different mechanism of secretion, and
are limited to axila and perianal area.

Sweat Secretion
Eccrine gland activity is regulated via neural
stimulation using sympathetic nerve fibers
distributed around the gland.
Neurotransmitter is acelylcholine
Sweating is controlled from hypothalamus (a
center in the preoptic and anterior regions),
where thermosensitive neurons are located.
The stimulus for perspiration:
direct heating alone (39 to 46oC)
physiological sweating due to nerve reflex arise from
sweat centers in brain cortex (emotional),
hypothalamus (thermoregulation)

Eccrine Sweat
contains mainly water (99.0 99.5%). It also
contains electrolytes NaCl, K+ and HCO3-, and
other simpl molecules - lactate, urea, ammonia,
amino acids (serin ornithin, citrulin, aspartic acid)
and minerals.
Mineral composition varies with the individual:

their acclimatisation to heat,

exercise and sweating,
the particular stress source (exercise, sauna, etc.),
the duration of sweating, and the composition of
minerals in the body

Apocrine Sweat
In lower mammals secretion of pheromones (trigger
sexual and territorial response)
In humans the significance of apocrine secretion of
pheromones is not completely understood.
Apocrine gland begin secreting at puberty
Apocrine duct exit to the surface via he hair follicle.
Apocrine sweat more viscous, with milky consistency
due to high content of fatty acids, cholesterol, squalene,
triglycerides, androgens, ammonia, sugars.

Mineral Composition of Sweat

sodium

0.9 g/l

potassium

0.2 g/l

calcium

15 mg/l

magnesium

1.3 mg/l

zinc

Microelements
0.4 mg/l

copper

0.3 0.8 mg/l

iron

1 mg/l

chromium

0.1 mg/l

nickel

0.05 mg/l

lead

0.05 mg/l

Sebaceous Glands
Glands secrete an oily/waxy
matter, called sebum, to
lubricate the skin and hair
Composition 25% wax
monoesters, 41% triglycerides,
16% free fatty acids, 1%
squalene, small amount
cholesterol esters and
cholesterol.

Skin Metabolism
Primary source for energy production in epidermis is glucose
from circulation diffuses into keratinocytes without effect of
insulin. Large proportion of glucose is catabolized up lactate
(even in presence of oxygen)
citric acid cycle does operate in epidermis explanation why this
cycle is inefficient is due to wide fluctuation of temperature and
blood flow in skin.

20% of glucose is metabolized by pentose-phosphate pathway

(PPP) production of NADPH and pentose for both FA
synthesis and nucleic acids.
Secondary source of energy - fatty acids derived from both
epidermal stores and exogenous sources (when glucose flow is
limited, then FA are metabolized).

Skin Metabolism
Glycogen small amount under physiological
conditions, however, elevation in all manner of injury of
epidermis or during hair growth in follicle explanation
energy when skin needs to be repaired or to use glucose
immediately, most probably disequilibrilium in
metabolic processes.
Furthermore, glucose is substrate also for synthesis of
lipids, polysaccharides, glycoproteins and nucleic acids.
GAG and proteoglycans highly charged and attract
water forming gels (see also lecture about ECM).

Skin Metabolism
Lipid metabolism - components:
a) membranes,
b) major constituents of permeability barrier,
c) energy supply
Synthesis from both glucose catabolism, from AA and
circulating FA - lipogenesis is going on in all layers of
epidermis - sebum synthesis in sebaceous glands
(higher synthesis of sebum is after sexual maturation).
Degradation - generally with lipases (yields in FA for
neutral lipids TG, sterol esters) in outermost layers of
epidermis (e.g. formation of prostaglandins)

Skin Immune System

Skin not only provides immune protection for itself, but
also for the whole body.
Cell types containing battery of mediators of immune
response
Langerhanse cells, monocytes, macrophages, mast cells
(cooperation with T-cells)

Cell types producing free radicals, anti-bacterial

peptides, cytokines chemokines, pro- and antiinflammatory mediators
Neutrophils, eosinophils, basophils.

B-cells secrete immunoglobulins (antibodies)