Krok-test

1. An electron micrograph shows a cell-to-cell

adhesion consisting, in each cell, of an attachment
plaque. The intercellular space is filled with
electron-dense substance including transmembrane
fibrous structures. What type of cytoskeleton
attached to plaque :
A. Microfilaments
B. Microtubules
C. Intermediate filaments
D. Actin filaments
E. Tubulin filaments
 Krok-test

2. The researchers destroyed the structure of

one of the cell parts during a scientific
experiment. As a result the cell lost the
ability to division. What structure was
broken?
A. Glycocalyx
B. Plastic complex
C. Microfibrille
D. Mitochondria
E. Centrosome
 Cytosol, inclusion,
translation

Melnyk P.20, 28-30

Ross, Pawlina P.31, 43-45, 70-71

Cytoplasm
Nucleus
 CYTOPLASM

Cytosol Inclusions
(hyaloplasm)

Cytosceleton Organelles
 The nuclear-cytoplasmic ratio (NC ratio)

The nuclear-cytoplasmic ratio (also

variously known as the nucleus:
cytoplasm ratio, nucleus-
cytoplasm ratio, N:C ratio, or N/C)
is a ratio of the size (i.e., volume)
of the nucleus of a cell to the size
of the cytoplasm of that cell.

NC = Vn / (Vc – Vn)
Vn – nuclear volume, Vc – cell volume.

NC ratio was proposed by Richard Hertwig in 1908. Richard Hertwig

 Water. Up to 90% in embryonic cells. 45-55% during aging
Proteins. About 20 % .
• enzymes that catalyze intermediate metabolism
reactions (that’s why cytosol is called an intermediate
cell metabolic system).
• proteins that cause processing (maturation) of
synthesized proteins,
• stressed proteins,
• chaperones,
• proteins of the elements of the cytoskeleton (actin,
tubulin), etc.
Lipids. For the most part, neutral fats, which form droplet
inclusions. Fatty acids and alcohols.
Carbohydrates. Mono-, di- and polysaccharides. Polysaccharides (starch in plant cells and
glycogen in animal), as well as neutral fats, form inclusion.
Ions. (Na+, K+, H+, Ca2+, Cl–, HCO3–, HPO42–, H2PO4–, HSO4–) etc.
Mineral (nutrient).
 The cytosol is also called hyaloplasm (from Greek
hyalos - glass) or cytoplasmic matrix, accounting
for about 55% of the total cell volume. It is a
complex colloidal cell system, it is internal
medium.

intermediate metabolism – a complex of chemical reactions for synthesis, decomposition and

modification of monomers of biopolymer molecules.
Glycolysis
Aggregate state of cytosol. It may vary depending on the action of the
environmental factors or the cell internal requires. It can be performed in two
aggregate states: sol (more liquid) and gel (thicker, gelatinous), that behaves like a
thixotropic gel. When cytosol is in the state of a gel, the particles of the dispersed
phase are interconnected into a three-dimensional grid. The dispersed medium is
contained in the cavities of this grid, thus, the whole system is deprived of fluidity.
When it is in a state of sol, then the three-dimensional grid is partially destroyed
and the system becomes more fluid. Moreover, its viscosity may vary in different
parts of the cell, which effects the reactions that occur there.
An important indicator of cytosol – pH, which is normally
about 6.8. The pH value in its various sections is
determined by introducing into the cell dye-indicators,
which change their color depending on the concentration
of hydrogen ions in the medium. The cytosol can act as a
buffer. So, if you add acids or alkalis to the intercellular
medium or enter them into a cell with a micro-pepper,
then the pH of the cytosol initially changes, but if the cell
is functioning normally, the pH of its cytosol is very
quickly restored to its level. Sustained pH maintenance is
required for the normal course of enzymatic reactions in
the cell.
1) It combines all cellular structures and ensures their chemical interaction.
2) All transport processes and processes of diffusion of various substances
take place here.
3) Osmotic and buffering properties of the cytoplasm are also mainly
provided by it.
4) In cytosol are localized enzymes involved in the synthesis and cleavage of
amino acids, nucleotides, fatty acids and carbohydrates, that is undergoing
intermediate metabolism reactions. Anaerobic cleavage of glucose -
glycolysis takes place, which results in the formation of two molecules of
ATP and tricarboxylic compounds.
5) In cytosol on free ribosomes or polysomes the synthesis of all protein in
the cell begins. Note that the synthesis of secretory proteins intended for
the extracellular matrix, the plasma membrane, the elements of the
vacuolar system, begins in cytosol, but ends on the membranes of the
granular endoplasmic reticulum, whereas the processes of protein
synthesis for the own needs of cytosol (enzymes for the synthesis and
cleavage of amino acids, nucleotides, carbohydrates , chaperones, etc.),
nucleus proteins, peroxisomes and a certain part of mitochondrial proteins
1) Proteins in the processes of broadcasting or immediately after its
completion must acquire the correct secondary, tertiary and
quaternary structure. The proper spatial organization of the protein
depends on its functional activity. Proteins that have an incorrectly
rolled polypeptide chain are functionally inactive. The process of
folding the polypeptide chain into the correct three-dimensional
structure is called maturation, or folding of proteins.
2) In addition, cytosol is a place where specific post-translational
modifications of proteins are carried out and under certain
conditions - their specific decomposition.
Translation
 tRNA
RNA transcription aminoacetilation
and processing

Nucleus Translation

Cytosol Posttranslation
modifications and
folding

Proteolysis
 acceptor area (for
attachment of amino acid)

anticodon

clover leaf

Nucleotide chain
Secondary (left) and tertiary (right) structure of tRNA

The function of tRNA is the delivery of amino acids to the ribosome for their
inclusion in the synthesized protein. The attachment of amino acids to tRNA
occurs clover leafin cytosol using the aminoacyl-tRNA synthetase enzyme.
Aminoacylation of
tRNA Required:
tRNA, amino
acid, Mg2 +,
ATP,
aminoacyl-
tRNA
synthetase
 Genetic code
The code defines how sequences of nucleotide triplets, called codons,
specify which amino acid will be added next during protein syntesis
 Translation

Stage I.
Initiation

Required:
ribosome, mRNA (capped with start codon AUG), initiation factors
(IF1, IF2, IF3 (in prokaryotes), eIF-2 (in eukaryotes), Mg2 +, GTP
 1.Codon- Stage II
depende
Elongation
nt
binding
Required: a
ribosomal complex
3.Translocation with mRNA and
peptide, GTP,
elongation factors
(eg EF-TU, EF-TS),
peptidyltransferase

2. Trans-
peptidation
Elongation
Elongation
 Stage III
Termination

Required:
a complex of ribosomes from
the mRNA (with one of the
stop codons (UAA, UAS,
UGA) and peptide, GTP,
release factors (termination
factors (for example, in
prokaryotes: RF1, RF2,
RF3, eRNA – in eukaryotes),
the factor that cleaves tRNA
Polysomes
Translation
mRNA Translation
Posttranslation modifications
 Unpolar side
Polar side groups groups
Folding
Folding is the process when a
synthesized polypeptide chain
Unfolded polypeptide obtains a definite working
conformation.

For folding of large proteins, are

Folded conformation in water medium required: chaperones and
foldases.

Hydrophobic part of
Polar side
core contains
groups can form
unpolar side groups
hydrogen bonds
Complexes of bacterial chaperones GroEL and GroES
 GroES/GroEL complex
(top)

Complexes of bacterial
chaperones GroEL and GroES

GroEL (side)
GroES/GroEL
complex (side)

GroEL (top)
Protein folding in the chaperone's cavity (Anfinsen's cavity)

GroEL
+ ATP

GroEL/ES

+ ATP

+ ADP + ATP
Mechanism of bacterial Chaperones GroES / GroEL complex action
 Chaperones and Prions

Polypeptide Polypeptide
PrP (Met) PrP (Val)

Chaperone Prion Chaperone Prion

PrP C PrP Sc PrP C The prion

(Prion) conformation is
not formed
 Creutzfeldt–Jakob disease (CJD)

Autopsy specimen from the frontal lobe of

patient with rapid cognitive decline,
myoclonic seizures, elevated 14-3-3
protein showing vacolated neuropil,
compatible with diagnosis of Creutzfeldt-
Jakob-Disease
Stanley B. Prusiner in 1997 obtained the Nobel Prize for discovering
prions
PrP C and PrP Sc
Formation of prions
Prions
 tRNA
RNA transcription aminoacetilation
and processing

Nucleus Translation

Cytosol Posttranslation
modifications and
folding

Proteolysis
 Lifetime of the protein molecule

Mechanisms of proteolysis of old proteins

For the first amino Ubiquitin-dependent

acid proteolysis

Amino acids
Met, Ser, Thr,
Ala, Val, Cys,
Gly and Pro -
stabilizing, the
rest -
destabilizing
26S-Proteasome
26S-Proteasome
Ubiquitin Proteasome System program
 Inclusions
Inclusion of the cytoplasm is an unnecessary cell component that appears and disappears
depending on the intensity and nature of the metabolism in the cell and on the conditions of
the organism's existence. Inclusions have the form of grains, debris, drops, vacuoles, granules
of various sizes and shapes. Their chemical nature is very diverse. Depending on the functional
purpose, inclusion devide into groups.
Classification by chemical nature
Inclusions
Glycosides
Proteins

Carbohydrates Terpenoids Alkaloids

Lipids Minerals
Classification by function
Inclusions

Trophic Secretory Excretory Pigmental Protective

 Trophic inclusions
These include fat drops, starch and aleiron grains, glycogen inclusion, yolk enzymes,
etc. Later, when the body is in unfavorable conditions, these stored substances are
used. Many trophic inclusions also contain animal egg cells and seed cells in plants.
These substances are used appropriately during embryonic development and
germination of seeds.

Beans seeds Seed germination

There are trophic lipid inclusions. They are in the form of droplets of different sizes found
both in plant and in animal cells. In animals they are contained in cells of white and brown
fatty tissue, as well as in liver cells. When starving, they split up with the release of a large
amount of energy stored in the form of ATP and used for energy needs of the cell. In cells of
brown fat tissue, when the fats break down, energy dissipated in the form of heat is
released in the cold season and goes to warm the body of the animal. Therefore, brown fat
tissue is most commonly found in animals that fall into winter hibernation. During the
decomposition of fats water is formed, so in some animals (for example, in camels) the
white fat tissue plays the role of water depot. Lipid inclusion can be detected using lipophilic
dyes (osmic acid, Sudan, etc.).

White fatty tissue Brown fatty tissue

Fat inclusion in liver cells
Adipocytes of Greater omentum
 Triglyceride
O
H2C O C (CH2) n CH3
O
HC O C (CH2) n CH3
O
H2C O C (CH2) n CH3

Hydrophobic part of
the molecule
Belong to trophic inclusions. By
chemical nature, they are blended
with protein-lipid. Contained in the
egg cells of almost all animals. They
are synthesized in the endoplasmic
reticulum, then modified and
segregated in the Golgi apparatus. The
yolk, synthesized by its own egg, is
called endogenous. In most living
organisms, yolk is synthesized not only
by the egg, but also by cells from
other organs, such as the liver, and
then goes to the egg. This is
exogenous yolk. The yolk inclusions
are used as energy and plastic
material in the development of the
embryo.

Human egg cell

 Yolk inclusions in amphibian blastomers
(containing proteins, lipids, RNA and phosphates)
 Glycogen

Such inclusions belong to carbohydrate trophic

inclusions of animal cells. Glycogen in its
chemical nature is a polysaccharide
constructed of glucose monomers. It is a white
powder, well soluble even in cold water. These
inclusions occur in cells of the liver, muscles. In
the case of inadequate nutrient intake,
glycogen decomposes into glucose molecules,
which are then included in the process of
glycolysis. At the same time the energy which
released is reserved in the form of ATP.

Detection of glycogen in the liver

Glycogen molecule by the method of Best.
Glycogen inclusions in liver cells. Carmin dying.
Hepatocyte
Glycogen inclusions
 Secretory inclusions
Secretory inclusions - various
substances of a chemical nature
(proteins, carbohydrates,
glycoproteins, etc.), which are
synthesized by the cell, extracted
from it and function outside of it.
They include secretory granules
with digestive enzymes in
pancreatic cells, as well as
hormones that "expect" an
appropriate signal to excrete from
the cell. Often, these inclusions
exist in the form of secretory
vesicles or bubbles surrounded by
a single membrane. Typically, such
vesicles are separated from the
Golgi complex, some time stay in
the cell and then by exocytosis Secretory granules in the pancreas
output their contents beyond the
cell.
Pancreacyte
Secretory granules
Hepatocytes
Endotheliocytes
Plasma membrane of the hepatocyte
Secretory vesicles containing protein
Excretory inclusions

Excretory inclusions - various

metabolic products that remain
in the cell for a specific time. An
example is the inclusion of
resins in coniferous plant cells.
In animal cells excretion
inclusions include the inclusion
of lipofuscin (aging pigment) in
neurons and some other cells
during aging.
 Pigmental inclusions
Melanin (from the Greek.
"melanos" - "black") belongs
to the pigmental and
protective inclusions.
Melanins are widespread in
plant tissues, animals and
even Protista. They
determine the color of the
skin and hair, such as the
horse's suit, the color of
feathers of birds (with
interference colors), fish
scales, insect cuticles. These
pigments also contain
iridescent eyes, sepia, and
the like. The inclusion of
Melanin in human skin
melanin is deposited in the
form of granules associated
with protein form
(melanoproteins).
Inclusion of melanin in pigmental skin cells
 Horse Colibri

Betta splendens Lucanus cervus

 The variety of human eye colors

Melanocytes of zebra skin

 Types of melanin
Melanin

Eumelanin Neuromelanin
Black and brown Pheomelanin In nervous system
Yellow

In humans during the growth of hair, the formation of pigments eumelanine and
pheomelanine occurs simultaneously. Thus, each hair, from the root to the tip, has
a color that depends on the combination of two of these pigments.
Black hair more than
99% consists of
eumelanine, chestnut
and light - about 95%
eumelanine and 5%
pheomelanine, red -
more than a third of
pheomelanine.
The color of the skin of people is due to constitutional pigmentation and depends on
the genetic prerequisites that provide for a diverse combination of the basic
pigments of the skin: melanin, carotene (yellow), oxyhemoglobin (red) and
deoxyhemoglobin (blue).
Black man and albino
Albinism is an inherited disease that is transmitted
from parents to children. The presence of genetic
disorders with albinism leads to the absence (or
blockade) of a special enzyme tyrosinase. Without it
it is impossible to synthesize melanin. As a rule, with
albinism there is a red eye, because reflected light
passes through the blood vessels in the iris, which
does not contain pigment.
Sepia - a complex nitrogen substance, the
secret of the ink gland (growth of the
rectum) of sea molluscs - cuttlefish, calamari.
Latin cuttlefish sounds Sepia, which is why
the secret of the mollusk was named. It
serves as a means of self-defense of an
animal, disorienting the enemy; When an
enemy collides with a "bomb", it explodes,
creating a "cloud of clouds". The secret of
one cuttlefish is able to color and make
opaque thousands of liters of water in a few
seconds. The secret of the ink is used in
painting as a very stable dark brown paint. Cuttlefish releases ink in water
The color of fresh sepia is almost black, but
at some time turns reddish brown. Natural
sepia is used in large quantities by the
industry for the manufacture of paint. The
paint comes from the English Channel and
the Mediterranean. Modern sepia is brought
from Sri Lanka. For the manufacture of sepia
ink bags of cephalopods are dried, finely
peeled, and the powder is boiled in a
solution of alkali. Using hydrochloric acid,
the dye is precipitated, washed with water
and dried at low temperature. Then he
thoroughly rubbed with rubber arabic and
goes on sale. Ink for sepia color pens
 Classification of pigments
Pigments

Carotenoids Quinones Flavonoids Derivatives of Other

porphyrin groups

The most widespread class of

biological pigments. They are
in all without exception plants
and many microorganisms.
Carotenoids cause the coloring
of many animals, especially
insects, birds and fish.
Carotenoids and their
derivatives are the basis of the
visual pigments responsible for
perception of light and color in
animals.
Lycopene is a non-cyclic isomer of beta-carotene. Protects parts of the plant from
sunlight and oxidative stress. In plant cells acts as a precursor to all other
carotenoids, including beta-carotene. It has a red color and breaks down fats.
Mainly contained in tomatoes, as well as in watermelon, pink grapefruit, and the
like.

Tomato (Solánum lycopérsicum) Watermelon (Citrullus)

Giovanni Stanchi «Watermelons, peaches, pears and other fruit in a landscape», 1645-1672
Pigments of this group contain a porphyrin complex. Gem, one of the species,
porphyrins, is part of the prosthetic group of hemoglobin, bilirubin, cytochrome
P450, and the like. This group also includes plant pigments - chlorophyll,
pheophytin. As a rule, pigments of this class are involved in photochemical
processes, as well as involved in the metabolism.
Hemoglobin
Hemoglobin (from the Greek haima - blood and lat. Globus - a bullet) is a complex
iron-binding protein of erythrocytes of animals and humans, capable of reversibly
binding to oxygen, ensuring its transfer to tissues.

The structure of the hemoglobin molecule Red blood cells

 Bilirubin
Bilirubin (Latin bilis - bile and ruber - red) - one
of the bile pigments. It is formed in the body of
blood hemoglobin; Isolated with bile in the
intestine. Excessive levels of bilirubin in the
blood occur in diseases that go beyond the
increased destruction of red blood cells or
disturbance of the excretory function of the
liver.
Crystals of bilirubin
Pigments, but NOT inclusions
Rhodopsin, or optic purple - the main visual
pigment. It is contained in the retinal sticks of
the marine invertebrates, fish, almost all
terrestrial vertebrates and humans. The
reconstructed rhodopsin (in low light) is
responsible for night vision, and at daylight,
the "colored dawn" (bright light) decomposes
and the maximum of its sensitivity shifts to the
Human sticks and cones under
blue area.
electron microscope

Activating rhodopsin with light Rhodopsin molecule

 Chlorophyll
Chlorophyll (from the Greek: chloros - "green" and phyllon - "leaf") - a green pigment,
present in cells of higher plants and some algae, giving them a corresponding color. It
helps plants get light energy. It is necessary for the process of photosynthesis.

Common features of the hema structure

and chlorophyll
Chlorella green algae
Pigments of plant leaves

leaf chloroplasts
 Cytochrome P450
The cytochrome P450 (cytochrome P450-dependent monooxygenase, CYP) is the
generic name of the P450 family of enzymes. Included in the class of hemoproteins
refer to cytochromes of type b. The cytochrome P450, bound to carbon monoxide,
has a maximum absorption of light at a wavelength of 450 nm, which determined its
name. Metabolism of many drugs occurs with the participation of cytochromes. In
addition, cytochromes provide the biosynthesis of steroids and cholesterol, the
metabolism of fatty acids and the metabolism of calcium.

Cytochrome P450 Fluorescent cytochrome P450

 Protective Inclusions
Protective inclusions are formed in cells of
plants and animals and serve to protect the
body. In animals, this type of inclusions can also
include melanin. With the help of melanin and
some other inclusions, a masking color is
created. So octopus and chameleon have the
ability to quickly change the intensity and
shades of its color precisely due to the
redistribution of pigments in the skin cells.
Octopus

Senegalese Chameleon Yemen chameleon

 Protective inclusion of plants

Triterpenes Steroids

Cardiotonic
Monoterpenes
Saponins

Flavonoids
Anthracenoglycosides

Plant glycosides Tannids

Thioglycosides (tannins)

Coumarins

Nitroglycosides
Phenolic glycosides Furanochromons

Phenolic Acids Phenol Spirits

 Monoterpene glycosides
Increase appetite, increase the secretion of the stomach and liver, peristalsis of the intestine.

Artemisia absinthium Centaurium

Menyanthes trifoliata Taraxacum

 Cardiotonic glycosides
Used for the treatment of heart failure and other heart diseases.

Strophanthus kombe

Adonis vernalis

Digitalis
 Triterpene glycosides (saponins)

́ glábra
Glycyrrhíza Polrmonium

Ginseng Equisetum arvense

 Saponins
Saponins (from the Latin Sapo - soup) - glycosides, the molecules of which are
formed by monosaccharides and terpenoid or steroid. Perform antiinflammatory,
antiallergic, tonic, anticoagulation, antispasmodic, stimulating, analgesic, choleretic,
sedative effects.

Steroid glycosides (saponins)

Dioscorea nipponica Echinopanax elatus

 Phenolol glycosides

Arctostáphylos

Rhodiola rusea
 Anthracene derivative glycosides
Phenolic compounds, derivatives of anthracene. Perform a mild, choleretic and
antimicrobial effects. Anthragliozides from Rubia tinctórum carry out lipolytic action.

Rubia tinctórum
Rúmex confértus Frángula álnus
 Coumarins and furanochromones

Ammi majus Pastináca sátiva

Peucedanum oreoselinum Psoralea drupacea

 Flavonoids
Phenolic compounds with two aromatic rings and a pyron cycle. Flavonoids perform
the secretory function of the stomach, antiinflammatory, reparative, diuretic,
hypoglycemic, antihypoxic, antispasmolithic, choleretic, sedative, hemopoietic
effects.

Tanacetum vulgare Leonurus cardiaca Origanum

 Tannids (tannins)
Polymeric phenols that perform antiinflammatory, astringent, antiseptic,
hemostatic effects. Used as antidote when poisoned with alkaloids.

Cotinus coggýgria Sanguisórba officinális

Thioglycosides

Brássica nígra
 Nitroglycosides
During enzymatic digestion in the gastrointestinal tract among the products of
decomposition they produce a hydrochloric acid (hydrogen cyanide), which is a
strong poison.

Prunus dulcis var. amara

The inclusion of alkaloids is in the cells of some plants. These are nitrogen-containing organic
heterocyclic compounds. The vast majority of alkaloids are toxic for humans and animals. They
are divided into two types: soluble and insoluble. Different plants contain different alkaloids:
nicotine - in tobacco, morphine - in poppy, quinine - in chinny wood, atropine - in belladonna,
caffeine - in coffee tree and tea bush, theobromine - in cocoa. In addition to the protective
function, they may be involved in the exchange of nitrogen-containing substances.

Papáver Theobroma cacao

Socrates Conīum maculātum

Alkaloid Coniine
 Ізопрен

Cinnamonum camphora Essential oils

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