QTransport of substances through cell membranes

Introduction
I. Characteristics of the plasma membrane
1. Composition
2. Lipids’ movement into the membrane
3. Selective permeability
II. Functions of the cell membrane proteins
1. Peripheral proteins
2. Integral proteins
a. Ligand-binding receptors
b. Adhesion molecules
c. Enzymes
d. Carriers, channels or pores
e. Intracellular signaling
3. Glycocalyx
III. Membrane transport
1. Gradient across the plasma membrane
2. Types of transport through the cell membranes 1
3. Passive transport
a. Simple diffusion or just diffusion
b. Facilitated diffusion
c. Osmosis
4. Active transport
a. Primary active transport
b. Secondary active transport
5. Transport through vesicles
a. Endocytosis
b. Exocytosis
c. Transcytosis
IV. Balance of fluids in the body
1. Daily intake of water
2. Daily loss of body water
a. Insensible water loss
b. Sweat
c. Water loss with feces
d. Water loss by the urine
3. Regulation of fluid exchange between intra and extracellular fluid
4. Hypertonic, isotonic, hypotonic fluids
5. Osmotic equilibrium
Physiology: Requires the study of the interaction of the molecules with the cells and the organs with the
organism.

Organization of the cell: Each cell is composed by the cytoplasm and in most of the cases the nucleus.
The nucleus is surrounded by the nuclear membrane and the cell by the plasma membrane or cellular
membrane.

Protoplasm : Includes all the matter in a cell, the cytoplasm and the nucleus

It contains the five types of substances that compose the cells

1. Water in the cells its concentration can reach 80% and many chemical substances are solved in it
2. Ions provide inorganic chemicals for cellular reactions and controls some of the cellular mechanisms
3. Carbohydrates main nutrient of the cells and is in the glycoproteins helping to identify our molecules
4. Lipids soluble in fat solvents have a big variety of properties
5. Proteins can be structural or functional proteins

Structural proteins
They are usually polymers of many individual protein molecules forming long filaments as:
1. microtubules of the “cytoskeletons” of organelles as cilia, nerve axons, the mitotic spindles of cells
undergoing mitosis, and a tangled mass of thin filamentous tubules that hold the parts of the cytoplasm
and nucleoplasm.
2. sarcomeres where muscle proteins facilitate contraction
3. Extracellularly, are in the connective tissue and in blood vessel walls, tendons, ligaments, and so forth
Functional Proteins : Functional proteins are mainly the enzymes of the cell. They often move in the cell
fluid, but also can bind to membranous structures inside the cell. Enzymes catalyze specific intracellular
chemical reactions reducing time, energy, and temperature.

Lipid’s functions :
- The fat stored in the adipocytes is the storehouse of energy-giving nutrients that can later be used 3
to provide energy.
- Cholesterol is responsible of the synthesis of hormones as Vit D and sexual hormones
- Send signals in the immune system and other cells
- Are used to form the cell membrane and intracellular membrane barriers that separate and
communicate the different cell compartments.
Membranes in the cell
Membranes in the cell surround different organelles and form:
a) cell membrane,
b) nuclear membrane,
c) membrane of the endoplasmic reticulum,
d) membranes of the mitochondria,
e) membranes of the lysosomes and other vesicles,
f) and the Golgi apparatus.

I. Characteristics of the plasma membrane

1. Composition

It is composed aprox:
- 55 % proteins
- 25% phospholipids
- 13% cholesterol
- 4% other lipids
- 3% carbohydrates
 Cells are water and the extracellular fluid too. To avoid mixing of intra and
extracellular compartments, cells have a bilayer of phospholipids and
other lipids that has the polar head next to water and a hydrophobic
structure blocking the water interchange in both sides of the membrane.

Because phospholipids combine hydrophilic heads with hydrophobic tails, their interaction with water is
referred to as amphipathic. And ions or water requires pores to be transported
Cholesterol confers certain rigidity to the membranes which lipids are constantly moving. Cholesterol
condenses lipids tails and forms ordered domains

2. Lipids’ movement into the membrane

The parallel sheets of phospholipids that face each other tail to tail are known as leaflets. At high
temperatures lateral diffusion can proceed rapidly, and the lipid is said to be in the sol state. At lower
temperatures, as the normal for humans, phospholipids diffuse slowly, and it is called the gel state.

Part of the lipids’ movements are controlled by proteins in the membrane: flippase and floppase, that
requires ATP; and scramblase, which is calcium dependent and flips one outer phopholipid to an inner
phospholipid leaflet, and also flips an inner phospholipid leaflet into an outer phospholipid leaflet
simultaneously.
 3. Selective permeability
It is selectively permeable to small molecules such as ions and metabolite. Protein molecules in the
membrane often penetrate all the way through the membrane, providing specialized pathways normally in
pores, for passage of specific substances through the membrane.

It must be able to accumulate substances against a concentration gradient. It is impermeable to large

molecules such as proteins and nucleic acids
Phospholipid bilayer membranes are impermeable to charged molecules. Phospholipid bilayer
membranes are impermeable to ions such as Na+, K+ , Cl- , and Ca2+. They cannot travel from one side of the
membrane to the opposite. The same is true of large water-soluble molecules, such as proteins, nucleic
acids, sugars, and nucleotides. They need carriers and proteins have that function
Small uncharged polar molecules can cross fairly freely the plasma membrane. This is often true for O2,
CO2, NH3 and for lipidic substances and molecules as alcohol. Water itself requires a carrier called
aquaporin.
 II. Functions of the cell membrane proteins
Integral proteins : They are intimately associated with the lipid bilayer. Three groups have been found:
transmembrane proteins that cross the bilayer; the embedded in the membrane without crossing it and
the lipid- anchored proteins with covalent bounds to a specific lipid molecule.
Peripheral proteins : They adhere tightly to the cytoplasmic or extracellular surfaces of the plasma
membrane
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1. Peripheral proteins
Mostly they are enzymes or work as controllers of transport of substances through the cell membrane
“pores.” They can participate as intracellular signaling. Also, they can bind to extracellular surface or to
integral proteins of the cytoskeleton.
 2. Integral proteins

a. Ligand-binding receptors

The interaction of a molecule with the extracellular portion of

protein initiates the transmission of the signal across the
membrane.

b. Adhesion molecules
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Form physical contacts with the surrounding extracellular matrix or with their cellular neighbors. • They

regulate the shape, growth, and differentiation of cells.

c. Enzymes
Many membrane-bound enzymes in the guts break down small polysaccharides or polypeptides into single
sugars or amino acids, for its absorption. This mechanism is repeated in numerous cell types, making the
membrane an efficient two-dimensional reaction center for multistep processes that involve enzymatic
reactions or transport.
Many of the GPI-linked proteins are enzymes. Several of the enzymatic activities that are classically
thought of as extracellular markers of the plasma membrane are attached to the external leaflet through
this lipid.
GPI group is composed by: - phosphatidylinositol group - glucosamine or mannose glycosidically bound to
the inositol of the phospholipid -ethanolamine phosphate that binds to the C-terminal amino acid of a
folded integrated enzyme

d. Carriers, channels or pores

For transporting substances that cannot cross the lipid bilayer.
Sometimes these carrier proteins transport substances in the direction opposite to their electrochemical
gradients for diffusion, which is called “active transport.” These pumps use the energy that is released
through the hydrolysis of ATP to drive the transport of substances into or out of cells against energy
gradients.
e. Intracellular signaling
They are all located at the intracellular leaflet of the membrane bilayer and often participate in intracellular signaling
pathways. Are linked to lipids. They include the small and heterotrimeric GTP-binding proteins, kinases, and
oncogene products. Many of these proteins are involved in relaying the signals that are received at the cell surface
to the effector machinery within the cell interior.
Translation for those who don't have a doctorate yet, lol :

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 3. Glycocalyx
Membrane carbohydrates are in combination with proteins or lipids in the form of glycoproteins or glycolipids. Most
of the integral proteins are glycoproteins, and about 1/10 of the membrane lipid molecules are glycolipids. The
“glyco” portions of these molecules almost invariably protrude to the outside of the cell, dangling outward from the
cell surface. The surface of the cell has a loose carbohydrate coat called the glycocalyx

Some carbohydrate substances bound to small protein cores, called proteoglycans are attached to the outer surface
of the cell as well. They are also included in the glycocalyx structure

Functions of the Glycocalyx :

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1. Has a negative electrical charge, which gives most cells an overall negative surface charge to repel other
negatively charged molecules.
2. Attach cells to one another.
3. Many of the carbohydrates act as receptor substances for binding hormones, and in turn, activate a
cascade of intracellular enzymes.
4. Some carbohydrate moieties enter into immune reactions, originating cell recognition proteins as the
Major Histocompatibility Complex
 III. Membrane transport

1. Gradient across the plasma membrane

There is a concentration gradient between exterior and the interior of the cells: outside there is a richer
concentration of some elements as O2, Na and Cl; inside, there is a higher concentration of K, CO2 and
proteins.
The cytosol has more negative charges (due to proteins) which produces an electrical gradient. It is called
membrane potential the difference in electric potential between the interior and the exterior of a
biological cell.

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Electrochemical gradient is the combined influence of the concentration of electrolytes and the electrical
gradients on the movement of each ion.
 2. Types of transport through the cell membranes

Movement of each molecule across the membrane in one direction or the other is known as unidirectional
flux. The algebraic sum of the two unidirectional fluxes is the net flux, or the net transport rate. Net
transport occurs only when the unidirectional fluxes are unequal.
Net transport takes place only when the net driving force is displaced from the equilibrium point.
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When no net driving force is acting on the substance is said that it is at equilibrium across the membrane.
But there may be equal and opposite movements of this molecule across the membrane.
Equilibrium is actually a special case of a steady state.
In a steady state, by definition, the conditions related to a substance do not change with time.
Thus, a transport system is in a steady state when both the driving forces acting on it and the rate of transport are
constant with time.

3. Passive transport

The substance goes from the higher concentration 14

area to the one with lower concentration. There
are 3 types : simple diffusion, facilitated diffusion,
osmosis

a. Simple diffusion or just diffusion

Factors that determine the rate

 The atom or molecule should be uncharged, hydrophobic to easily cross the lipid bilayer.
 The speed of kinetic motion of the substance.
 The number and diameter of the virtual spaces in the membrane. Molecular weight cut off.
  The properties of the membrane are very relevant:
- The difference composition of lipids in the bilayer will change the permeability
- Also the thickness of the membrane and the surface that allows the flux of the molecule
- Those will determine the permeability coefficient of each type of membrane
- All those variable are included in Flick’s law: flux=permeability coefficient X difference in
concentration

In simple diffusion, the flux of an uncharged substance through membrane lipid is directly proportional to
its concentration difference 15
The rate of uptake reach a maximum in facilitated diffusion, because the carriers are saturated if the
concentration rise.

b. Facilitated diffusion through protein channels

 Lipid-insoluble molecules can pass through the protein pore
channels
Many of the body’s cell membranes contain protein “pores”
called aquaporins that selectively permit rapid passage of
water through the membrane. There are at least 13 different
types in cells.

Protein channels are distinguished by:

- They are selectively permeable to certain substances,
- Many of the channels can be opened or closed by gates that are regulated by electrical signals
(voltage gated channels) or chemicals that bind to the channel proteins (ligand-gated channels).

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Selective permeability of protein channels :

Many of the protein channels are highly selective for transport of specific ions or molecules. The selectivity
is due to the characteristics of the channel: diameter, shape, the nature of the electrical charges and
chemical bonds along its inside surfaces. For example, in the K channels, at the top of the channel pore are
pore loops that form a narrow selectivity filter. Lining the selectivity filter are carbonyl oxygens.

Gating of protein channels :

The opening and closing of gates are controlled in 2 ways:

1. Voltage gating. responds to the electrical potential across the cell membrane. This process is
the basic mechanism for eliciting action potentials in nerves.
 2. Chemical (ligand) gating. Some protein channel gates are opened by the binding of a chemical
substance (a ligand) with the protein, which causes a conformational or chemical bonding
change in the protein molecule that opens or closes the gate.
We have also :
3. Mechanically gated. Responds to pressure
4. Always open

Carrier proteins :

Carriers bind the substance and have a conformational change to transport it to the other part of the
membrane. They get saturated and the rate of transport at that point becomes constant. Their conduit 17
never offers a continuous transmembrane path because it has at least two gates that are never open at the
same time

c. Osmosis

By far the most abundant substance that diffuses through the cell membrane is water. Normally, the
volume of the cell remains constant. However, under certain conditions, a concentration difference for
water can develop across a membrane is called osmosis. Water moves to equalize the molarity in both
sides of the membrane, that is semipermeable
Osmolarity and osmotic pressure:

Osmolarity is the osmolar concentration expressed as osmoles per liter of solution rather than osmoles per
kilogram of water

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Strictly speaking, it is osmoles per kilogram of water (osmolality) that determines osmotic pressure, but for
dilute solutions, the quantitative differences between osmolarity and osmolality are less than 1 percent.
The amount of pressure required to stop osmosis is called the osmotic pressure. Then, should be applied
against the flow of water.

Types of solutions
Normally in our body all the solutions are isotonic (same osmolarity). Sometimes, a solution can be diluted
by an excess of water (hypotonic solution) or concentrated by an excess of whichever molecule (hypertonic
solution).

Clinical consequences

Clinically, osmolarity problems are caused by decreases in [Na+ ] in plasma and ECF(extra cellular fluid)
(hyponatremia), increases in [Na+ ] (hypernatremia), or increases in glucose concentration
(hyperglycemia). Changes in the concentration of urea, which accumulates in kidney failure, have no
effect on tonicity.

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6. Active transport
Active transport is a process that can transfer a solute through the membrane against its electrochemical
potential energy difference. • Requires energy and a protein to facilitate the transport. • It is used by many
ions (K+, Na+ , Ca2+, H+ ,Cl-, Fe3+), amino acids, some sugars, etc.
There are two types:
1. Primary Active transport 2. Secondary Active transport
 a. Primary active transport
Na-K pump :
The driving force needed to cause net transfer of a solute against its electrochemical gradient comes from
the favorable energy change that is associated with an exergonic chemical reaction, such as ATP hydrolysis.
The prototype is Na-K pump
Their functions: a) control the cellular volume b) maintain a negative electrical voltage

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With each forward cycle, the pump couples the extrusion of three Na+ ions and the uptake of two K+ ions
to the intracellular hydrolysis of one ATP molecule. By themselves, the transport steps of the Na-K pump
are energetically uphill; that is, if the pump were not an ATPase, the transporter would run in reverse. In
cells with high Na-K pump rates, such as renal proximal tubules, a third or more of cellular energy
metabolism is devoted to supplying ATP to the Na-K pump.
Calcium pump
Calcium are at low concentration in the intracellular cytosol.
There are two primary active transport calcium pumps:
1. In the cell membrane and pumps calcium to the outside of the cell. 21
2. In the membrane of one or more of the intracellular vesicular organelles of the cell (endoplasmic
reticulum)
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The SERCAs (for sarcoplasmic and endoplasmic

reticulum calcium ATPases) appear to transport two H+
and two Ca2+ ions for each molecule of ATP hydrolyzed.

Protons pumps
Primary active transport of hydrogen ions is important in 2 organs:
1) in the gastric glands of the stomach.
2) in the late distal tubules and cortical collecting ducts of the kidneys.
But other types are present in - Mitochondria - Intracellular organelles as lysosomes or Golgi apparatus
 b. Secondary active transport
• Includes two types :
- Cotransporters or symporters
- Counter-transport or exchangers

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Cotransporters or symporters :
Carry two or more solutes in the same direction
The gradient of Na represents a storehouse of energy because the excess sodium outside the cell
membrane is always attempting to diffuse to the interior. This serves as a storehouse of energy
The carrier then serves as an attachment point for both the sodium ion and the substance to be co-
transported. Once they both attached, the energy gradient of Na causes both substances to be transported
together to the cytoplasm.

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The co-transported substance could be glucose or an amino acid. • Cotransporters are present in epithelial
cells of the intestines and renal tubules.

Exchangers or counter-transport
They move two or more molecules in opposite directions.

Sodium-calcium counter-transport occurs in most of our cells; sodium-hydrogen counter-transport is found in the
proximal renal tubules; and chloride-bicarbonate exchanger located in neurons, the intercalated cells of the
collecting ducts of the kidneys and the erythrocytes, colon, lung and breast cells
The Na energy diffusing into the cell allows other substance to be transported to the outside. Na binds to
the carrier protein outside, while the substance to be countertransported binds to the interior part of the
protein. Then a conformational change occurs, and the Na energy takes out the other substance.

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7. Transport through vesicles

Evaginations or invaginations of the plasma and organelles

membranes are used to transport substances that cannot
cross the membranes.
Those include:
- endocytosis pinocytosis or bulk-phase endocytosis
phagocytosis
- exocytosis
- and transcytosis.

a. Endocytosis
Endocytosis is the process of capturing a substance or particle from outside the cell by engulfing it with the
cell membrane. The membrane folds over the substance and it becomes completely enclosed by the
membrane. At this point a membrane-bound sac, or vesicle, pinches off and moves the substance into the
cytosol.
According with the content of the vesicle, are distinguished two types:
- phagocytosis
- pinocytosis

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Phagocytosis :

Phagocytosis, or cellular eating, occurs when solid molecules enter the

cell. The plasma membrane engulfs the solid material, forming a
phagocytic vesicle with pseudopodia. Normally phagocytosis has a
defensive purpose

Pynocytosis :
 Pinocytosis is a type of endocytosis in which small particles
suspended in the extracellular fluid are moved into the cell
through pores formed on the cell membrane. The content is
liquid and the substances are taken by invagination. The main
purpose is the incorporation of molecules to the cell. In
humans, pinocytosis is mostly associated with the absorption
of fat.

Although involves the binding of molecules to receptors in the

membrane, forming coated vesicles, it is different from other
receptormediated endocytosis processes as the receptor is not
specific for one molecule.

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Receptor-mediated endocytosis :

Receptor-mediated endocytosis is a highly selective type of endocytosis by which cells take up specific
ligands. The budding of transport vesicles and the selective incorporation of cargo into the forming vesicles
are both mediated by protein coats which are supramolecular assemblies of proteins recruited from the
cytosol.

b. Exocytosis
Exocytosis is the process of vesicles fusing with the plasma membrane and releasing their contents to the outside of
the cell. It exports substances, such as a protein, or helps the cell to get rid of a waste product or a toxin. Newly
made membrane protein and membrane lipids are moved on top the plasma membrane by exocytosis

Receptor-mediated endocitosis and exocytosis :

The coats of proteins deform flat membrane patches into round buds, eventually leading to the release of
coated transport vesicles. The coats also participate in cargo selection by recognizing sorting signals
present in the cytosolic domains of transmembrane cargo proteins. The first coats to be identified and
characterized contained a scaffold protein, clathrin, as their main constituent

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Now have been discovered no-clathrin coats that contains proteins as - COPII that mediates transport to
the Endoplasmic Reticulum to the Golgi apparatus - or the SNARE complex responsible of the fusion of
membranes of vesicles for exocytosis and assures the specificity of transport

c. Transcytosis
Is the transfer of molecules across cells from one side to the other, a process that entails endocytosis,
vesicular transfer and exocytosis, and which speeds the bulk movement of molecules through tissues. In
some cases, transcytosis is receptor-mediated, and is often carried out by vesicles called caveolae
 V. Balance of fluids in the body
The relative constancy of the body fluids is remarkable because there is continuous exchange of fluid and
solutes with the external environment, as well as within the different body compartments. Fluid intake is
highly variable and must be carefully matched by equal output of water from the body to prevent body
fluid volumes from increasing or decreasing.

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1. Daily intake of water

Water is added to the body by two major sources: (1) it is ingested in the form of liquids or water in food,
which together normally add about 2100 ml/day to the body fluids, and (2) it is synthesized in the body by
oxidation of carbohydrates, adding about 200 ml/day. • These mechanisms provide a total water intake of
about 2300 ml/day
Intake of water is highly variable among different people and even within the same person on different
days, depending on climate, habits, and level of physical activity

2. Daily loss of body water

There are 4 types of losses of water:

- insensible water loss
- fluid loss in sweat
- water loss in feces
- water loss by urine
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a. Insensible water loss

Humans experience a continuous loss of water by evaporation from the respiratory tract and diffusion
through the skin, which together account for about 700 ml/day of water loss under normal conditions. •
This loss is termed insensible water loss because we are not consciously aware of it, even though it occurs
continually in all living humans.
Insensible water loss through the skin occurs is different of sweating and is present even in people without
sweat glands; the average water loss by diffusion through the skin is about 300 to 400 ml/day.
This loss is minimized by the cholesterol-filled cornified layer of the skin, which provides a barrier against
excessive loss by diffusion. When the cornified layer becomes denuded, the rate of evaporation can
increase as much as 10-fold, to 3 to 5 L/day.

Insensible water loss through the respiratory tract averages about 300 to 400 ml/day. As air enters the respiratory
tract, it becomes saturated with moisture, to a vapor pressure of about 47 mm Hg, before it is expelled.

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The vapor pressure of the inspired air is usually less than 47 mm Hg, water is continuously lost through the
lungs with respiration. In cold weather, the atmospheric vapor pressure decreases to nearly 0, causing a
greater loss of water from the lungs as the temperature decreases.
b. Sweat
The amount of water lost by sweating is highly variable, depending on physical activity and environmental
temperature. The volume of sweat normally is about 100 ml/day, but in very hot weather or during heavy
exercise fluid loss in sweat occasionally increases to 1 to 2 L/hour.
c. Water loss with feces
Only a small amount of water (100 ml/day) is lost in the feces. This loss can increase to several liters a day
in people with severe diarrhea. For this reason, severe diarrhea can be life threatening if not corrected
within a few days
 d. Water loss by the urine
Urine represents the biggest amount of excreted liquid. The rate of urine excretion is highly controlled to
maintain the balance between water and electrolytes intake and output. Urine volume can be as low as 0.5
L/day in a dehydrated person or as high as several L/day in a person who has been drinking tremendous
amounts of water.

This variability of intake is also true for most of the electrolytes of the body, such as Na, Cl and K. The daily
sodium intake may vary from 20 to 300 mEq/day. The kidneys adjust the excretion rate of water and
electrolytes to match precisely the intake of these substances, as well as compensating for excessive losses
of fluids and electrolytes that occur in certain disease states

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3. Regulation of fluid exchange between intra and extracellular fluid

The relative amounts of extracellular fluid distributed between the plasma and interstitial spaces are
determined mainly by the balance of hydrostatic and colloid osmotic forces across the capillary
membranes.
The distribution of fluid between intracellular and extracellular compartments, is determined mainly by the
osmotic effect especially Na, Cl, and other electrolytes acting across the cell membrane, giving that the cell
membranes are highly permeable to water (due to aquaporins) but relatively impermeable to even small
ions such. Water moves across the cell membrane rapidly and both sides of the membrane remains
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isotonic.

4. Hypertonic, isotonic, hypotonic fluids

A isotonic solution neither shrinks nor swells the cells, because it has the same osmolarity than the cells.
Solutions prescribed to patients, as physiological serum (0,9% NaCl or 5% glucose) can be infused into the
blood without upsetting osmotic equilibrium between the intracellular and extracellular fluids. Solutions
with a higher concentration of impermeant solutes, are hypertonic and will shrink the cells. Solutions with
a lower concentration of impermeant solutes, are hypotonic and will swell the cells.
 5. Osmotic equilibrium
Osmotic equilibrium is rapidly attained between the intra and extracellular spaces. The transfer of fluid
across the plasma membrane in culture occurs rapidly correcting differences in osmolarities within seconds
or minutes. But complete equilibrium the whole body takes about 30 minutes usually after drinking water,
because the liquid should be absorbed and transported to all the tissues.

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