Exercise 1: Cell Transport and Permeability

Overview:
Plasma membrane
its molecular composition allows it to be selective
about what passes through it
allows nutrients and appropriate amounts of ions to
enter the cell and keeps out undesirable substances
selectively permeable: valuable cell proteins and
other substances are kept w/in the cell; metabolic
wastes pass to the exterior
transport through it occurs in 2 basic ways:
Passive process - driven by concentration or
pressure differences (gradients) bet the interior
and exterior of the cell
Active process- cell provides energy (ATP) to

power the transport

Diffusion and filtration: 2 key passive processes of
membrane transport
Simple diffusion - occurs w/o the assistance of
membrane proteins; diffusion of solute particles
dissolved in water through a selectively
permeable membrane
Facilitated diffusion - requires membranebound carrier protein that assists in the transport

In both simple and facilitated diffusion, the substance

being transported moves w/ (along or down) the
concentration gradient of the solute (from a region of
its higher conc to a region of its lower conc)

The process does not require energy from the

cell
Energy in the form of kinetic energy comes
from the constant motion of the molecules

Movement of solutes continues until they are

evenly dispersed throughout the solution - reached
equilibrium
Osmosis - special type of diffusion where water
moves w/ its concentration gradient, from a higher
conc of water to a lower conc of water (diffusion of
water
through
a
differentially
permeable
membrane)
It moves in response to a higher conc of
solutes on the other side of a membrane

Filtration - usually occurs only across capillary

walls in the body
Depends upon a pressure gradient as its
driving force
Not a selective process
Dependent upon the size of the pores in the filter

Active transport and passive transport: 2 key

active process
Active transport - uses a membrane-bound
carrier protein like f. diffusion but differs from it
bec the solutes move against their conc gradient
and bec ATP is used to power the transport
Vesicular transport - includes phago, endo,
pino, and exocytosis

Act 1: Simulating Dialysis (Simple Diffusion)

Intro
When a conc gradient (difference in conc) exists, the
net effect of random molecular movement of molecules
is that they become evenly distributed throughout the
environment - diffusion occurs
Kinetic energy of the molecules is the driving force
behind diffusion
All molecules possess KE and are in constant
motion
As molecules move about randomly at high
speeds, they collide and bounce off one another,
changing direction w/ each collision
For a given temp, all matter has about the same
average kinetic energy
Smaller molecules tend to moves faster than larger
mol bec KE is directly related to both mass and
velocity (KE = 1/2 mv2)
The diffusion of particles into and out of cells is modified
by the plasma membrane w/c is a physical barrier
In general, molecules diffuse passively through the
plasma membrane if they are small enough to pass
through its pores (and are aided by an electrical
and/or conc gradient) or if they can dissolve in the
lipid portion of the membrane (as in the case of O 2
and CO2)
A membrane is selectively permeable/ differentially

permeable/ semipermeable if it allows some

particles to pass but not others

*The dialysis membranes used each have a different

molecular weight cutoff (MWCO), indicated by the
num below it
*Think of MWCO in terms of pore size
*The larger the MWCO num, the larger the pores in
the membrane
*Molecular weight of a solute is the number of grams
per mole where mole is the constant Avogadros num
6x1023 molecules/mole
*The larger the molecular weight, the larger the mass of
the molecule
*Molecular mass is sometimes used instead if molecular
weight

Experiment:
- 20 MWCO membrane to the membrane holder
- increase NaCl conc to be dispensed to the left beaker to
9.00mM
- deionized water - right beaker
- after starting the run, the barrier bet the beakers will
descend, allowing the solutions in each beaker to have
access to the dialysis membranes separating them
- you will be able to determine the amt of solute that
passes through the membrane by observing the conc
display to the side of each beaker
- a level above 0 in NaCl conc in the right beaker
indicates that Na and Cl ions are diffusing from the left
beaker into the right through the selectively permeable
dialysis membrane
- NaCl: no diffusion

The reason sodium chloride didn't diffuse left to right

is that the membrane pore size was too small.
- flush each beaker for next run
The molecular weight of urea is 60.07. Do you think
urea will diffuse through the 20 MWCO membrane? No, not at all.
- increase urea conc to to be dispensed to the left
beaker to 9.00 mM; deionized - right, dispense, run,
flush
- urea: no diffusion
Glucose is a six-carbon sugar. Albumin is a protein
with 607 amino acids. The average molecular weight
of a single amino acid is 135 g/mole. There is no

reason to run these solutes at the 20 MWCO

because glucose and albumin are both too large to
pass.
- 50 MWCO
- increase NaCl to 9.00 mM (left); deionized - right
- NaCl: equilibrium at 10 min
- increase NaCl conc to 18mM (left) ; deionized right
- NaCl: equilibrium at 10 min
- 100 MWCO
- NaCl to left 9.00 mM; deionized right
- NaCl: equilibrium at 10 min
- increase urea conc to 9.00 mM (left); deionized right
- urea: equilibrium at 16 min
The rate of diffusion for urea is slower than that for
sodium because urea is a larger molecule.
Recall that glucose is a monosaccharide, albumin
is a protein with 607 amino acids, and the average
molecular weight of a single amino acid is 135
g/mole. Which of the following will be able to
diffuse through the 200 MWCO membrane? glucose only
- 200 MWCO
- increase glucose to 9.00 mM (left); deionized right
- glucose: equil at 37 min
- increase albumin to 9.00 mM (left); deionized -

right
- albumin: no diffusion
Pre-lab Quiz Results
1. The driving force for diffusion is
- the kinetic energy of the molecules in motion.
2. In diffusion, molecules move
- from high concentration to low concentration.
3. Which of the following dialysis membranes has the
largest pore size?
- 200 MWCO
4. Avogadro's number is a constant for the number of
- molecules
Post-lab Quiz Results
1. The effect of increasing the concentration of sodium
chloride from 9 mM to 18 mM in the left beaker was to
- increase the rate of diffusion
2. Describe the difference between the rate of diffusion
seen for sodium and urea.
- Urea diffused more slowly because it is larger than
sodium
3. Which of the following solutes did not pass through
any of the membranes?
- albumin
4. When diffusion stops, we say the solution has reached
- equilibrium
Review Sheet Results
1. Describe two variables that affect the rate of diffusion.
- size and concentration of solute
2. Why do you think the urea was not able to diffuse
through the 20 MWCO membrane? How well did the
results compare with your prediction?
- As predicted, Urea was too big to pass through the 20
MWCO membrane
3. Describe the results of the attempts to diffuse glucose
and albumin through the 200 MWCO membrane. How
well did the results compare with your prediction?
- The glucose diffused through the 200MWCO
membrane, while there was no diffusion of albumin as
predicted. Albumin is too big to pass through 200
MWCO membrane
4. Put the following in order from smallest to largest
molecular weight: glucose, sodium chloride, albumin,
and urea.
- Sodium chloride, Urea, Glucose, Albumin
Activity 2: Simulated Facilitated Diffusion

rate of facilitated diffusion

Intro
Some molecules are lipid insoluble or too large too
pass through the pores in the cells plasma
membrane so they pass via facilitated diffusion
Example: sugars, amino acids, ions
Solutes combine w/ carrier-protein molecules in
the membrane and are then transported w/ their
conc gradient
The carrier-protein molecules in the membrane
might have to change shape slightly to
accommodate the solute but the cell does not
have to expend the energy of ATP
Solute transport varies w/ the num of available
carrier-protein molecules in the membrane
Carrier proteins can become saturated if too
much solute is present and the maximum
transport is reached
Carrier proteins are embedded in the plasma
membrane and act like a shield, protecting the
hydrophilic solute from the lipid portions of the
membrane
Facilitated diffusion typically occurs in one
direction for a given solute
The greater the conc difference bet one side
of the membrane and the other, the grater the

Experiment:
- note: glucose carriers display in the membrane builder is
set at 500
- build membrane to insert 500 glucose carrier proteins into
the membrane then drag to membrane holder
- increase glucose conc to be dispensed to the left beaker
to 2.00 mM; deionized - right
- after starting the run, the barrier bet the beakers will
descend, allowing the solutions in each beaker to have
access to the dialysis membranes separating them
- you will be able to determine the amt of solute that
passes through the membrane by observing the conc
display to the side of each beaker

- a level above 0 in glucose conc in the right beaker

indicates that glucose is diffusing from the left beaker
into the right through the selectively permeable dialysis
membrane
- glucose: equil at 43 min
- increase glucose conc to 8.00mM (left)
- glucose: equil at 58 min

What happened to the glucose transport rate when

the glucose concentration was increased from 2
mM to
8 mM? - The glucose transport rate increased
- return to membrane builder, dispense
What effect do you think increasing the number of
protein carriers will have on the glucose transport
rate? - The glucose transport rate will increase

- increase the num of glucose carriers to 700

- increase glucose conc to 2.00 mM (left)
- glucose: equil at 33 min
- increase glucose conc to 8.00 mM (left)
- glucose: equil at 43 min

Describe the conditions that resulted in the fastest rate

of glucose transport. - 700 glucose carrier proteins and
8 mM glucose
- decrease the num of glucose carriers to 100
- increase glucose conc to 10.00 mM (left)
- glucose: equil not reached
Why do you think equilibrium wasn't reached? - The
membrane proteins were saturated
- increase num of glucose carriers to 700
What effect do you think adding Na+ Cl- will have on
the glucose transport rate? - There will be no change in
the glucose transport rate
- increase glucose conc to 2.00 mM (left)
- increase NaCl conc to be dispensed to the right beaker
to 2.00 mM
- NaCl: equil at 8 min
- glucose: equil at 33 min
Pre-lab Quiz Results
1. Molecules need a carrier protein to help them move
across a membrane because
- they are lipid insoluble or they are too large
2. Which of the following is true of facilitated diffusion?
- Movement is passive and down a concentration
gradient
3. Examples of solutes that might require facilitated
diffusion include
- all of the above
4. Which of the following would not affect the rate of
facilitated diffusion?
- the amount of intracellular ATP
Post-lab Quiz Results
1. What variable(s) increased the rate of facilitated
diffusion of glucose?
- both a and b
2. For facilitated diffusion, increasing the concentration
of glucose on one side of the membrane is the same as
- making the concentration gradient steeper
3. When all of the membrane carriers are engaged, or
busy, we say they are
- saturated
4. Na+ Cl- had no effect on glucose transport because
- sodium is not required for glucose transport in the
simulation.

Review Sheet Results

1. Explain one way in which facilitated diffusion is the
same as simple diffusion and one way in which it is
different from
simple diffusion.
- Both of diffusion are passive diffusion that are from
high concentration to a low concentration. Facilitated
diffusion differs from simple diffusion because
facilitated diffusion requires a carrier protein to
diffuse
2. The larger value obtained when more glucose
carriers were present corresponds to an increase in the
rate of glucose
transport. Explain why the rate increased. How well
did the results compare with your prediction?
- There is more way for glucose. So rate of
diffusion was increased, as predicted
3. Explain your prediction for the effect Na+ Clmight have on glucose transport. In other words,
explain why you picked the choice that you did.
How well did the results compare with your
prediction?
- The rate of glucose diffusion did not change
because sodium chloride does not use protein
carrier so it does not help or hinder glucose
diffusion
Activity 3: Simulating Osmotic Pressure


Intro
Because water can pass through the pores of most
membranes, it can move from one side of a membrane
to the other relatively freely
Osmosis takes place whenever there is a difference in
water conc bet the 2 sides of a membrane
If we place distilled water on both sides of a
membrane, net movement of water does not occur,
however, water molecules would still move bet the 2
sides of the membrane (no net osmosis)
The conc of water in a solution depends on the num of
solute particles present
Increasing the solute conc coincides w/
decreasing the water conc
Bec water moves down the conc gradient
(higher to lower conc), it always moves toward
the solution w/ the highest conc of solutes
Solutes also moves down the their conc gradient
If we position a fully permeable membrane
(permeable to solutes and water) bet 2 solutions of
differing conc, then all substances (solutes and water)
diffuse freely and an equilibrium will be reached bet
the 2 sides of the membrane
However, if we use a selectively permeable
membrane that is impermeable to the solutes, then
we have established a condition where water moves
but solutes do not
Consequently, water moves toward the more
concentrated solution, resulting in a vol increase on
that side of the membrane

By applying this concept to a closed system where

volumes cant change, we can predict that the
pressure in the more conc solution will rise
Osmotic pressure - force that would need to be
applied to oppose the osmosis in a closed system

Osmotic pressure is measured in millimeters of mercury

(mm Hg)
In general, the more impermeable the solutes, the
higher the osmotic pressure
Osmotic changes can affect the vol of a cell when it is
placed in various solutions
Tonicity - the way a solution affects the vol of a cell
Tells us whether or not a cell will shrink or swell
Isotonic - if the conc of impermeable solutes is the
same inside and outside the cell
Hypertonic - higher conc of impermeable solutes
outside the cell than in the cells interior
- cell would shrink bec net movement of water
would be out of the cell

Hypotonic - conc of impermeable solutes is

lower outside the cell than in the interior
- cell would swell and probably burst bec net
movement of water would be into the cell

Experiment:
- 20 MWCO
- increase the NaCl conc to be dispensed to the left beaker
to 5.00 mM; deionized - right
- after starting the run, the barrier bet the beakers will
descend, allowing the solutions in each beaker to have
access to the dialysis membranes separating them
- you will be able to determine the changes in pressure in
the 2 beakers

- also the amt of solute that passes through the membrane by

observing the conc display to the side of each beaker
- a level above 0 in NaCl conc in the right beaker indicates
that Na and Cl are diffusing from the left beaker into the right
through the selectively permeable dialysis membrane
- NaCl: equil not reached
- osmotic equil reached
- increase NaCl conc to 10.00 mM
- NaCl: equil not reached
- osmotic equil reached

What effect do you think increasing the Na+ Clconcentration will have? - increased pressure
- 50 MWCO
- increase NaCl conc to 10.00 mM (left)
- NaCl: equil at 10 min

Why do you think there is no pressure change? - Sodium

is able to diffuse through the pores
- 100 MWCO
- increase glucose conc to 8.00 mM (left)
- glucose: no diffusion
- osmotic equil reached
- increase glucose conc to be dispensed to the right
beaker to 8.00 mM
- glucose: no diffusion
Why do you think there was no pressure change? There is no net movement of water because the solute
concentrations are the same
- 200 MWCO
- increase glucose conc to 8.00 mM (left)

- glucose: equil at 37 min

- increase albumin to 9.00 mM (left)
- increase glucose to 10.00 mM (right)
- albumin: no diffusion
- glucose: equil at 38 min
- osmotic equil reached
What do you think will be the pressure result of the
current experimental conditions? - pressure above
the left beaker
Act 3: Simulating Filtration

Intro
Filtration - process by w/c water and soultes pass
through a membrane (such as a dialysis
membrane) from an area of hydrostatic (fluid)
pressure into an area of lower hydrostatic pressure
Ex: fluids and solutes filter out of the capillaries in
the kidneys into the kidney tubules bec blood
pressure in the capillaries is greater than the fluid
pressure in the tubules. If blood pressure increases,
rate of filt also increases
The amt of filtrate (fluids and solutes that pass
through a membrane) depends almost entirely in
the pressure gradient (difference in pressure bet the
solutions on the 2 sides of the membrane) and on

the size of the membrane pores

Solutes that are too large to pass through are retained
by the capillaries, these usually include blood cells and
proteins
Ions and smaller molecules, such as glucose and
urea, can pass through

*In this act, the pore size is measured as a molecular

weight cutoff (MWCO), indicated by the num below it
*Think of MWCO in terms of pore size
*The larger the MWCO num, the larger the pores in the
membrane
*Molecular weight of a solute is the number of grams per
mole where mole is the constant Avogadros num 6x10 23
molecules/mole

*You will also analyze the filt membrane for the presence or
absence of solutes that might be sticking to the membrane

- 20 MWCO
- increase conc of NaCl, urea, glucose, and powdered
charcoal to be dispensed to 5.00 mM
- after starting the run, the membrane holder below the top
beaker retracts, and the solution will filter through the membrane
into the beaker below
- you will be able to determine whether solute particles are
moving through the filt membrane
- a rise in detected solute conc indicates that the solute particles
are moving through the filt membrane
- note: pressure is set at 50 mm Hg and the timer is set to 60
mins

- filtration stopped
- projected completion at 100 min
- drag the 20 MWCO membrane to the holder in the
membrane residue analysis unit

The reason none of the solutes were present in the

filtrate was that the solutes were all too large to pass
through.
What effect will increasing the pore size of the filter
have on the filtration rate? - The filtration rate will
increase
- 50 MWCO
- same conc of NaCl, urea, and glucose
- filt completed in 40 min
- analysis

- 200 MWCO
- same conc
- filt completed in 10 min
- analysis
What will happen if you increase the pressure above
the beaker (the driving pressure)? - The filtration rate
will increase
- increase the pressure to 100 mm Hg
- 200 MWCO
- same conc
- filt completed in 5 mins
What does an increase in the driving pressure
correspond to in the body? - an increase in blood
pressure

Act 5: Simulating Active Transport

Intro
Substances moved across cell membranes by an active
transport are generally unable to pass by diffusion
Reasons why a substance might not be able to pass
through a membrane by diffusion: might be too large
to pass through the pores, might not be lipid soluble,
might have to move against rather than w/ a
concentration gradient
The type of active transport wherein the substances
move across the membrane by combining w/ a carrierprotein
molecule
resembles
enzyme-substrate
interaction
ATP hydrolysis provides the driving force and the
substance move against conc gradients or
electrochemical gradients or both
Solute pumps - carrier proteins
Substances that are moved into cells by solute pumps
include amino acids and some sugars; both are
necessary for the life of the cell but they are lipid
insoluble and are too large too pass through the
membrane pores
Sodium ions (Na) are ejected from the cells by active
transport
There is more Na outside the cell than inside the
cell so Na tends to remain in the cell unless

actively transported out

In the body, the most common type of solute pump
is the Na-K (sodium-potassium) pump w/c moves
Na and K in opposite directions across cellular

membranes
3 Na ions are ejected from the cell for every 2
K ions entering the cell
There is more K inside the cell than outside
the cell so K tends to remain outside the cell
unless actively transported in
Membrane carrier proteins that move more than
one substance such as the Na-K pump, participate
in coupled transport
Carrier is a symporter if the solutes move in the
same direction
Carrier is antiporter if the solutes move in opposite
directions
Uniporter - carrier that transports only a single
solute

Experiment:
- note: number of Na-K pumps is set at 500
- click dispense to the left of the beaker to deliver 9.00 mM
NaCl solution to the cell
- increase KCl conc to be delivered to the beaker to 6.00
mM
- click dispense to the right of the beaker to deliver 6.00
mM KCl solution to the beaker
- increase ATP conc to 1.00 mM
- dispense ATP to deliver 1.00 mM ATP to both sides of
membrane
- after starting the run, the solutes will move across the cell
membrane, simulating active transport
- you will be able to determine the amt of solute that is
transported across the membrane
- NaCl: transport finished at 3 min
- KCl: transport finished at 3 min
- flush

Why did the sodium transport stop before the transport

was completed? - The ATP was depleted
- click dispense to the left of the beaker to deliver 9.00 mM
NaCl solution to the cell
- increase KCl conc to be delivered to the beaker to 6.00 mM
- click dispense to the right of the beaker to deliver 6.00 mM
KCl solution to the beaker
- increase ATP conc to 3.00 mM
- dispense ATP to deliver 3.00 mM ATP to both sides of
membrane
- NaCl: transport finished at 30 min
- KCl: transport finished at 30 min

- flush
- click dispense to the left of the beaker to deliver 9.00 mM
NaCl solution to the cell
- click deionized water to the right of the beaker and then
click dispense to deliver deionized water to the beaker
- increase ATP conc to 3.00 mM
- dispense ATP to deliver 3.00 mM ATP to both sides of
membrane

- NaCl: no transport
What do you think will result from these experimental
conditions? - No Na+ will be transported
- increase the num of Na-K pumps to 800
- click dispense to the left of the beaker to deliver 9.00 mM
NaCl solution to the cell
- increase KCl conc to be delivered to the beaker to 6.00
mM
- click dispense to the right of the beaker to deliver 6.00
mM KCl solution to the beaker
- increase ATP conc to 3.00 mM
- dispense ATP to deliver 3.00 mM ATP to both sides of
membrane

- NaCl: transport finished at 18 min

- KCl: transport finished at 18 min
Why was the equilibrium for the solutes reached
earlier? - There were more pumps for transport
- W/ the num of Na-K pumps still set to 800, increase
the num of glucose carriers t0 400
- click dispense to the left of the beaker to deliver 9.00 mM
NaCl solution to the cell

Do you think the addition of glucose carriers will

affect the transport of sodium or potassium? - No, it will

not affect the transport of either ion
- increase the KCl conc to be delivered to the beaker to
6.00 mM
- increase the glucose conc to be delivered to the beaker
to 10.00 mM
- click dispense to the right of the beaker to deliver 6.00 mM
KCl solution to the beaker and 10.00 mM glucose solution to
the beaker
- increase ATP conc to 3.00 mM
- dispense ATP to deliver 3.00 mM ATP to both sides of
membrane

- NaCl: transport finished at 18 min

- KCl: transport finished at 18 min
- Glucose: transport interrupted, not finished

Pre-lab Quiz Results

1. The movement of sodium and potassium maintained
by the Na+ -K+ pump
- requires energy and is against a concentration
gradient
2. The sodium-potassium pump is classified as a(n)
- antiporter
3. The sodium-potassium pump moves _____ sodium
ions and _____ potassium ions simultaneously.
- 3, 2
4. Solutes that require active transport for movement
might be too large to pass or might be
- lipid insoluble
Post-lab Quiz Results
1. What happened when you increased the amount of
ATP dispensed with the same concentration of sodium
and
potassium on either side of the membrane?
- More ions were transported
2. At what concentration of ATP were the sodium and
potassium maximally transported?
- 3 mM ATP
3. What was the effect of adding more Na+ -K+
pumps to the simulated cell?
- Transport of the ions was faster
4. Describe the effect of adding glucose carriers to the
sodium and potassium transport.
- There was no change in the transport rate because
glucose is transported independently
Review Sheet Results

1. Describe the significance of using 9 mM sodium

chloride inside the cell and 6 mM potassium
chloride outside the cell,
instead of other concentration ratios.
- The Na+/K+ pump works with 3:2 ratio of sodium
- potassium
2. Explain why there was no sodium transport even
though ATP was present. How well did the results
compare with your prediction?
- In order to observe working of Na+/K+ pump
sodium and potassium need to be present. So
sodium alone can not make pump work.
3. Explain why the addition of glucose carriers had
no effect on sodium or potassium transport. How
well did the results
compare with your prediction?
- Addition of Glucose and its carriers do not help or
hinder Na+/K+ pumping. Because glucose transport
works independently
4. Do you think glucose is being actively
transported or transported by facilitated diffusion in
this experiment? Explain your answer.
- It may transported by facilitated diffusion. Cause
glucose is a lipid insoluble and too large to pass
through the cell
membrane, it requires a glucose carrier