Transport

mechanisms
Lecture 10
Most essential learning
competencies
• Describe the structural components of the cell membrane
• Relate the structure and composition of the cell membrane
to its function
• Explain transport mechanisms in cells (diffusion, osmosis,
facilitated transport, active transport)
• Differentiate exocytosis and endocytosis
Cell membrane
An introduction
Cell membrane
• Also termed, plasma membrane
• Boundary that separates the
living cell from its
surroundings
• Controls traffic into and
out of the cell
• Gate keeper system
• Exhibits selective
permeability
Cell membranes are fluid mosaics of
lipids and proteins
Why is it called fluid mosaic
model?
• The membrane is fluid in structure with a “mosaic”
of various proteins embedded in or attached to a
double layer (bilayer) of phospholipids
Phospholipid
• A phospholipid is an
amphipathic molecule,
meaning it has both a
hydrophilic (“water-
loving”) region and a
hydrophobic (“water-
fearing”) region
Phospholipid
The proteins embedded in the
bilayer
• Like membrane lipids, most membrane proteins are
amphipathic. Such proteins can reside in the
phospholipid bilayer with their hydrophilic regions
protruding.
Phospholipids and Proteins
• This molecular orientation maximizes contact of
hydrophilic regions of a protein with water in the
cytosol and extracellular fluid, while providing
their hydrophobic parts with a nonaqueous
environment.
Membrane fluidity
• Refers to the movement of phospholipids
• Phospholipids are held together by hydrophobic
reactions, which are much weaker covalent bonds
• Movement of phospholipids: lateral, flip-flop or
transverse
Membrane fluidity
Membrane fluidity
• A membrane remains fluid as temperature decreases
until the phospholipids settle into a closely
packed arrangement and the membrane solidifies,
much as bacon grease forms lard when it cools.
• The temperature at which a membrane solidifies
depends on the types of lipids it is made of.
Unsaturated vs saturated
hydrocarbon tails
• As the temperature decreases, the membrane remains
fluid to a lower temperature if it is rich in
phospholipids with unsaturated hydrocarbon tails
• Because of kinks in the tails where double bonds
are located, unsaturated hydrocarbon tails cannot
pack together as closely as saturated hydrocarbon
tails, making the membrane more fluid
Cholesterol within the animal
cell membrane
• The steroid cholesterol, which is wedged between
phospholipid molecules in the plasma membranes of
animal cells, has different effects on membrane
fluidity at different temperatures.
• At relatively high temperatures— at 37°C, the body
temperature of humans, for example— cholesterol
makes the membrane less fluid by restraining
phospholipid movement.
Cholesterol within the animal
cell membrane
• However, because cholesterol also hinders the close
packing of phospholipids, it lowers the temperature
required for the membrane to solidify.
• Thus, cholesterol can be thought of as a “fluidity
buffer” for the membrane, resisting changes in
membrane fluidity that can be caused by changes in
temperature.
Plant cholesterol
• Compared to animals, plants have very low levels of
cholesterol; rather, related steroid lipids buffer
membrane fluidity in plant cells
Membranes must be fluid to work
properly
• The fluidity of a membrane affects both its
permeability and the ability of membrane proteins
to move to where their function is needed.
• When a membrane solidifies, its permeability
changes, and enzymatic proteins in the membrane may
become inactive if their activity requires movement
within the membrane.
Challenges in membrane fluidity
• However, membranes that are too fluid cannot
support protein function either.
• Therefore, extreme environments (for example, those
with extreme temperatures) pose a challenge for
life, resulting in evolutionary adaptations that
include differences in membrane lipid composition.
Membrane proteins and their
function
Two major populations of membrane proteins:
• Integral proteins
• penetrate the hydrophobic interior of the lipid bilayer
• The majority are transmembrane proteins, which span the
membrane
• Peripheral proteins
• are not embedded in the lipid bilayer at all; they are
loosely bound to the surface of the membrane, often to
exposed parts of integral proteins
Some functions of membrane
proteins
• Transport
• Enzymatic activity
• Signal transduction
• Cell-cell recognition
• Intercellular joining
• Attachment to the cytoskeleton and extracellular
matrix (ECM)
Simple Diffusion
Part 1
Simple Diffusion
• The movement of particles of any substance down to
its own concentration gradient
• The diffusion of a substance across a biological
membrane is called passive transport because it
requires no energy.
Simple Diffusion
• In the process of diffusion, a substance tends to
move from an area of high concentration to an area
of low concentration until its concentration
becomes equal throughout a space.
For example, think about someone opening a
bottle of cleaning ammonia in the middle of
a room. The ammonia molecules will initially
be most concentrated right where the person
opened the bottle, with few or no molecules
at the edges of the room.

Gradually, the ammonia molecules will

diffuse, or spread, away from the place
where they were released, and eventually
you’ll be able to smell ammonia at the edges
of the room. Ultimately, if the bottle is
capped and the room is closed, the ammonia
molecules will become evenly distributed
throughout its volume.
To understand this, imagine
that there’s an area where
molecules are more concentrated
(such as where ammonia has just
been opened) and an area where
they’re less concentrated (the
surrounding room).

Since there are lots of ammonia

molecules in the concentrated
area, it’s pretty likely that
one will move from there into
the non-concentrated area. But
since there are few molecules
of ammonia in the non-
concentrated area, it’s pretty
unlikely that the reverse will
happen.
Thus, over time, the net movement of molecules will
be out of the more concentrated area and into the
less concentrated one, until the concentrations
become equal (at which point, it’s equally likely
for a molecule to move in either direction).
This process does not require any energy input; in
fact, a concentration gradient itself is a form of
stored (potential) energy, and this energy is used
up as the concentrations equalize.
 Molecules can move
through the cell’s
cytosol by
diffusion, and some
molecules also
diffuse across the
plasma membrane.

Each individual substance in a solution or space has its

own concentration gradient, independent of the
concentration gradients of other materials, and will
diffuse according to that gradient.
Other factors being equal, a stronger concentration
gradient (larger concentration difference between
regions) results in faster diffusion.

Thus, in a single cell, there can be different rates

and directions of diffusion for different
molecules.

For example, oxygen might move into the cell by

diffusion, while at the same time, carbon dioxide
might move out in obedience to its own concentration
gradient.
Must watch: Cell - Diffusion

• https://www.youtube.com/watch?v=71MSBEwMGDA
Diffusion and osmosis
• Osmosis is a specific type of diffusion; it is the
passage of water from a region of high water
concentration through a semi-permeable membrane to
a region of low water concentration.
Osmosis
• Water solutions are very important in biology.
• When water is mixed with other molecules this
mixture is called a solution.
• Water is the solvent and the dissolved substance is
the solute.
• A solution is characterized by the solute. For
example, water and sugar would be characterized as
a sugar solution.
Osmosis
• The classic example used to demonstrate osmosis and
osmotic pressure is to immerse red blood cells into
sugar solutions of various concentrations.
• There are three possible relationships that cells
can encounter when placed into a sugar solution.
Isotonic solution
• The concentration of solute in the solution can
be equal to the concentration of solute in cells.
In this situation the cell is in an isotonic
solution (iso = equal or the same as normal). A red
blood cell will retain its normal shape in this
environment as the amount of water entering the
cell is the same as the amount leaving the cell.
Hypertonic solution
• The concentration of solute in the solution can
be greater than the concentration of solute in the
cells. This cell is described as being in
a hypertonic solution (hyper = greater than
normal). In this situation, a red blood will appear
to shrink as the water flows out of the cell and
into the surrounding environment.
Hypotonic solution
• The concentration of solute in the solution can
be less than the concentration of solute in the
cells. This cell is in a hypotonic solution (hypo =
less than normal). A red blood cell in this
environment will become visibly swollen and
potentially rupture as water rushes into the cell.
Must watch
• Osmosis
• Part 1 -
https://www.youtube.com/watch?v=SD1AKWUazPU
• Part 2 -
https://www.youtube.com/watch?v=MCvbfqz7ASs
Facilitated Transport
Part 2
Facilitated diffusion
• Some molecules, such as carbon dioxide and oxygen,
can diffuse across the plasma membrane directly,
but others need help to cross its hydrophobic core.

• In facilitated diffusion, molecules diffuse across

the plasma membrane with assistance from membrane
proteins, such as channels and carriers.
Facilitated diffusion
• A concentration gradient exists for these
molecules, so they have the potential to diffuse
into (or out of) the cell by moving down it.
However, because they are charged or polar, they
can't cross the phospholipid part of the membrane
without help.
Facilitated transport proteins
• Facilitated transport proteins shield these
molecules from the hydrophobic core of the
membrane, providing a route by which they can
cross.
• Two major classes of facilitated transport proteins
are channels and carrier proteins.
Channel proteins
• Channel proteins span the membrane and make
hydrophilic tunnels across it, allowing their
target molecules to pass through by diffusion.
• Channels are very selective and will accept only
one type of molecule (or a few closely related
molecules) for transport.
• Passage through a channel protein allows polar and
charged compounds to avoid the hydrophobic core of
the plasma membrane, which would otherwise slow or
block their entry into the cell.
Channel proteins
Aquaporins are channel
proteins that allow
water to cross the
membrane very quickly,
and they play
important roles in
plant cells, red blood
cells, and certain
parts of the kidney
(where they minimize
the amount of water
lost as urine).
• Some channel proteins are open all the time, but others
are “gated,” meaning that the channel can open or close
in response to a particular signal (like an electrical
signal or the binding of a molecule).

• Cells involved in the transmission of electrical

signals, such as nerve and muscle cells, have gated ion
channels for sodium, potassium, and calcium ions in
their membranes.

• The opening and closing of these channels, and the

resulting shifts in ion levels inside the cell, play an
important role in electrical transmission along
membranes (in nerve cells) and in muscle contraction
(in muscle cells).
Carrier proteins
• Carrier proteins can
change their shape to
move a target
molecule from one
side of the membrane
to the other.
Carrier proteins
• Like channel proteins, carrier proteins are typically
selective for one or a few substances.

• Often, they will change shape in response to binding of

their target molecule, with the shape change moving the
molecule to the opposite side of the membrane.

• The carrier proteins involved in facilitated diffusion

simply provide hydrophilic molecules with a way to move
down an existing concentration gradient (rather than
acting as pumps).
Facilitated transport proteins
• Channel and carrier proteins transport material at
different rates.
• In general, channel proteins transport molecules much
more quickly than do carrier proteins.
• This is because channel proteins are simple tunnels;
unlike carrier proteins, they don’t need to change
shape and “reset” each time they move a molecule.
• A typical channel protein might facilitate diffusion at
a rate of tens of millions of molecules per second,
whereas a carrier protein might work at a rate of a
thousand or so molecules per second
Must watch
• Facilitated diffusion -
https://www.youtube.com/watch?v=9aEaqJieb7g
Active Transport
Part 3
Why switch to active transport?
Passive transport is a great strategy for moving
molecules into or out of a cell. It's cheap, it's
easy, and all the cell has to do is sit there and
let the molecules diffuse in. But...it also doesn't
work in every situation.
For instance, suppose the sugar glucose is more
concentrated inside of a cell than outside. If the
cell needs more sugar in to meet its metabolic
needs, how can it get that sugar in?
Introducing active transport
• Here, the cell can't import glucose for free using
diffusion, because the natural tendency of the
glucose will be to diffuse out rather than
flowing in.
• Instead, the cell must bring in more glucose
molecules via active transport. In active
transport, unlike passive transport, the cell
expends energy (for example, in the form of ATP) to
move a substance against its concentration
gradient.
What is are these electrochemical
gradients?
• Atoms and molecules can form ions and carry
positive or negative electrical charges, there may
also be an electrical gradient, or difference in
charge, across a plasma membrane.
• In fact, living cells typically have what’s called
a membrane potential, an electrical potential
difference (voltage) across their cell membrane.
• An electrical potential difference exists whenever
there is a net separation of charges in space.

• In the case of a cell, positive and negative charges

are separated by the barrier of the cell membrane, with
the inside of the cell having extra negative charges
relative to the outside.

• The membrane potential of a typical cell is -40 to -80

millivolts, with the minus sign meaning that inside of
the cell is more negative than the outside

• The cell actively maintains this membrane potential,

and we’ll see how it forms in the section on the
sodium-potassium pump.
• As an example of how the
membrane potential can
affect ion movement, let’s
look at sodium and
potassium ions.

• In general, the inside of

a cell has a higher
concentration of potassium
(K+) and a lower
concentration of sodium
(Na+) than the
extracellular fluid around
it.
• If sodium ions are outside of a cell, they
will tend to move into the cell based on
both their concentration gradient (the
lower concentration of Na+) and the
voltage across the membrane (the more
negative charge on the inside of the
membrane).

• Because K+ is positive, the voltage across

the membrane will encourage its movement
into the cell, but its concentration
gradient will tend to drive it out of the
cell (towards the region of lower
concentration). The final concentrations
of potassium on the two sides of the
membrane will be a balance between these
opposing forces.

• The combination of concentration gradient

and voltage that affects an ion’s movement
is called the electrochemical gradient.
Active transport: moving against
a gradient
• To move substances against a concentration or
electrochemical gradient, a cell must use
energy.
• Active transport mechanisms do just this,
expending energy (often in the form of ATP)
to maintain the right concentrations of ions
and molecules in living cells.
• In fact, cells spend much of the energy they
harvest in metabolism to keep their active
transport processes running.
• For instance, most of a red blood cell’s
energy is used to maintain internal sodium
and potassium levels that differ from those
of the surrounding environment.
Active transport mechanisms can
be divided into two categories.
• Primary active transport directly uses a source of
chemical energy (e.g., ATP) to move molecules
across a membrane against their gradient.
• Secondary active transport (cotransport), on the
other hand, uses an electrochemical gradient –
generated by active transport – as an energy source
to move molecules against their gradient, and thus
does not directly require a chemical source of
energy such as ATP.
Primary active transport
• One of the most important pumps in animal cells is
the sodium-potassium pump, which moves Na+ out of
cells, and K+ into them.
• Because the transport process uses ATP as an energy
source, it is considered an example of primary
active transport.
• Not only does the sodium-potassium pump maintain
correct concentrations of Na+ and K+ in living
cells, but it also plays a major role in generating
the voltage across the cell membrane in animal
cells.
• Pumps like this, which are involved in the
establishment and maintenance of membrane voltages,
are known as electrogenic pumps.
• The primary electrogenic pump in plants is one that
pumps hydrogen ions (H+) rather than sodium and
potassium
The sodium-potassium pump cycle
The sodium-potassium pump
The sodium-potassium pump transports sodium out of
and potassium into the cell in a repeating cycle of
conformational (shape) changes.
In each cycle, three sodium ions exit the cell,
while two potassium ions enter.
This process takes place in the
following steps:
1. To begin, the pump is open to the inside of the cell. In
this form, the pump really likes to bind (has a high
affinity for) sodium ions, and will take up three of them.
2. When the sodium ions bind, they trigger the pump to
hydrolyze (break down) ATP. One phosphate group from ATP
is attached to the pump, which is then said to be
phosphorylated. ADP is released as a by-product.
3. Phosphorylation makes the pump change shape, re-orienting
itself so it opens towards the extracellular space. In
this conformation, the pump no longer likes to bind to
sodium ions (has a low affinity for them), so the three
sodium ions are released outside the cell.
3. In its outward-facing form, the pump switches
allegiances and now really likes to bind to (has a
high affinity for) potassium ions. It will bind
two of them, and this triggers removal of the
phosphate group attached to the pump in step 2.
4. With the phosphate group gone, the pump will
change back to its original form, opening towards
the interior of the cell.
5. In its inward-facing shape, the pump loses its
interest in (has a low affinity for) potassium
ions, so the two potassium ions will be released
into the cytoplasm. The pump is now back to where
it was in step 1, and the cycle can begin again.
This may seem like a complicated cycle, but it just
involves the protein going back and forth between
two forms: an inward-facing form with high affinity
for sodium (and low affinity for potassium) and an
outward-facing form with high affinity for potassium
(and low affinity for sodium).
The protein can be toggled back and forth between
these forms by the addition or removal of a
phosphate group, which is in turn controlled by the
binding of the ions to be transported.
Secondary active transport
• The electrochemical gradients set up by primary active
transport store energy, which can be released as the
ions move back down their gradients.
• Secondary active transport uses the energy stored in
these gradients to move other substances against their
own gradients.
• As an example, let's suppose we have a high
concentration of sodium ions in the extracellular space
(thanks to the hard work of the sodium-potassium pump).
If a route such as a channel or carrier protein is
open, sodium ions will move down their concentration
gradient and return to the interior of the cell.
Secondary active transport
• In secondary active transport, the movement of the
sodium ions down their gradient is coupled to the
uphill transport of other substances by a shared
carrier protein (a cotransporter).
• For instance, in the figure below, a carrier
protein lets sodium ions move down their gradient,
but simultaneously brings a glucose molecule up its
gradient and into the cell. The carrier protein
uses the energy of the sodium gradient to drive the
transport of glucose molecules.
Secondary active transport
• In secondary active transport, the two molecules
being transported may move either in the same
direction (i.e., both into the cell), or in
opposite directions (i.e., one into and one out of
the cell). When they move in the same direction,
the protein that transports them is called
a symporter, while if they move in opposite
directions, the protein is called an antiporter.
Secondary active transport
Must watch
• Primary vs secondary transport -
https://www.youtube.com/watch?v=N-iBdwtQn4Q
• Active Transport – primary and secondary -
https://www.youtube.com/watch?v=xmXCswxPjQg
 NOTICE:
Endocytosis will be the topic
for reporting instead of bulk
transport.

Exocytosis will be the topic for

reporting instead of vesicular
transport.
Bulk Transport
• Macromolecules are too large to move with membrane
proteins and must be transported across membranes
in vesicles
• Two types of bulk transport: Endocytosis and
Exocytosis.
Endocytosis
Part 4
Endocytosis
• Endocytosis is a type of active transport that
moves particles, such as large molecules, parts of
cells, and even whole cells, into a cell.
• There are different variations of endocytosis, but
all share a common characteristic: the plasma
membrane of the cell invaginates, forming a pocket
around the target particle.
• The pocket pinches off, resulting in the particle
being contained in a newly created intracellular
vesicle formed from the plasma membrane.
• There are three types of endocytosis: Phagocytosis,
pinocytosis and receptor-mediated endocytosis.
Phagocytosis
• Phagocytosis (the condition of “cell eating”) is
the process by which large particles, such as cells
or relatively large particles, are taken in by a
cell.
• For example, when microorganisms invade the human
body, a type of white blood cell called a
neutrophil will remove the invaders through this
process, surrounding and engulfing the
microorganism, which is then destroyed by the
neutrophil (Figure 1).
• In preparation for phagocytosis, a portion of the inward-facing
surface of the plasma membrane becomes coated with a protein
called clathrin, which stabilizes this section of the membrane.

• The coated portion of the membrane then extends from the body of
the cell and surrounds the particle, eventually enclosing it.

• Once the vesicle containing the particle is enclosed within the

cell, the clathrin disengages from the membrane and the vesicle
merges with a lysosome for the breakdown of the material in the
newly formed compartment (endosome).

• When accessible nutrients from the degradation of the vesicular

contents have been extracted, the newly formed endosome merges
with the plasma membrane and releases its contents into the
extracellular fluid.

• The endosomal membrane again becomes part of the plasma membrane.

Pinocytosis
• A variation of endocytosis is called pinocytosis.
This literally means “cell drinking” and was named
at a time when the assumption was that the cell was
purposefully taking in extracellular fluid.
• In reality, this is a process that takes in
molecules, including water, which the cell needs
from the extracellular fluid.
• Pinocytosis results in a much smaller vesicle than
does phagocytosis, and the vesicle does not need to
merge with a lysosome (Figure 2).
• A variation of pinocytosis is called potocytosis. This
process uses a coating protein, called caveolin, on the
cytoplasmic side of the plasma membrane, which performs
a similar function to clathrin.

• The cavities in the plasma membrane that form the

vacuoles have membrane receptors and lipid rafts in
addition to caveolin.

• The vacuoles or vesicles formed in caveolae (singular

caveola) are smaller than those in pinocytosis.

• Potocytosis is used to bring small molecules into the

cell and to transport these molecules through the cell
for their release on the other side of the cell, a
process called transcytosis.
Receptor-Mediated
Endocytosis
• A targeted variation of
endocytosis employs
receptor proteins in the
plasma membrane that
have a specific binding
affinity for certain
substances (Figure 3).
• In receptor-mediated endocytosis, as in
phagocytosis, clathrin is attached to the
cytoplasmic side of the plasma membrane.

• If uptake of a compound is dependent on receptor-

mediated endocytosis and the process is
ineffective, the material will not be removed from
the tissue fluids or blood.

• Instead, it will stay in those fluids and increase

in concentration.
• Some human diseases are caused by the failure of
receptor-mediated endocytosis.

• For example, the form of cholesterol termed low-density

lipoprotein or LDL (also referred to as “bad”
cholesterol) is removed from the blood by receptor-
mediated endocytosis.

• In the human genetic disease familial

hypercholesterolemia, the LDL receptors are defective
or missing entirely. People with this condition have
life-threatening levels of cholesterol in their blood,
because their cells cannot clear LDL particles from
their blood.
• Although receptor-mediated endocytosis is designed
to bring specific substances that are normally
found in the extracellular fluid into the cell,
other substances may gain entry into the cell at
the same site.

• Flu viruses, diphtheria, and cholera toxin all have

sites that cross-react with normal receptor-binding
sites and gain entry into cells.
Must watch
• Endocytosis -
https://www.youtube.com/watch?v=QspmZf_yWyU
• Receptor-mediated endocytosis -
https://www.youtube.com/watch?v=PjrH1dpCyyE
Exocytosis
Part 5
Exocytosis
• The reverse process of moving material into a cell
is the process of exocytosis.
• Exocytosis is the opposite of the processes
discussed in the last section in that its purpose
is to expel material from the cell into the
extracellular fluid.
• Waste material is enveloped in a membrane and fuses
with the interior of the plasma membrane.
• This fusion opens the membranous envelope on the
exterior of the cell, and the waste material is
expelled into the extracellular space (Figure 4).
• Other examples of cells releasing molecules via
exocytosis include the secretion of proteins of the
extracellular matrix and secretion of
neurotransmitters into the synaptic cleft by
synaptic vesicles.
Exocytosis
• Cells must take in certain molecules, such as
nutrients, but they also need to release other
molecules, such as signaling proteins and waste
products, to the outside environment.
• Exocytosis (exo = external, cytosis = transport
mechanism) is a form of bulk transport in which
materials are transported from the inside to the
outside of the cell in membrane-bound vesicles that
fuse with the plasma membrane.
• Some of these vesicles come from the Golgi
apparatus and contain proteins made specifically by
the cell for release outside, such as signaling
molecules.
• Other vesicles contain wastes that the cell needs
to dispose of, such as the leftovers that remain
after a phagocytosed particle has been digested.
• These vesicles are transported to the edge of the
cell, where they can fuse with the plasma membrane
and release their contents into the extracellular
space.
• Some vesicles fuse completely with the membrane and
are incorporated into it, while others follow the
“kiss-and-run” model, fusing just enough to release
their contents (“kissing” the membrane) before
pinching off again and returning to the cell
interior
Two types of Exocytosis
• Constitutive secretion is the default pathway and
is used primarily to replenish material at the
plasma membrane and certain membrane-bound
organelles.
• Regulated secretion terminates in secretory
vesicles that store secreted material until a
signal triggers fusion with the plasma membrane.