Chapter 4: Transport of substances
through cell membranes
Cell membrane = lipid bilayer + transport
proteins (penetrate bilayer)
Lipid bilayer = immiscible with extra/
intracellular fluid: acts as a barrier against
movement of water/ water-soluble m.
Lipid soluble m. diffuse directly thru lipid
substance.
Mem proteins: alternate pathway thru
mem: penetrating proteins: mainly
transport proteins:
• Channel proteins: watery spaces thru
m., allow free movement of water,
selected ions/m.
• Carrier proteins: bind with
molecules/ ions to be transported →
conformational change → move
substance to other side of mem.
• Usually both types are selective
Diffusion
Random molecular movement of
substances m. by m. thru intermolecular
spaces in mem/ carrier protein.
Energy causing diffusion = normal Kinetic
motion of matter (constant motion = ‘heat’)
Greater motion, higher temperature.
Motion stops at absolute zero temperature.
A approaches stationary B: electrostatic,
other nuclear forces of A repel B,
transferring some KE of A → B: random
continual movement of m. among one
another in liquids/ gases = diffusion
Suspended colloid particles diffuse in
similarly to ions/ whole m. but slower due
to large size.
Diffusion thru cell mem:
• Simple: Kinetic movement of m./ ions
occurs thru mem opening/
intermolecular spaces without
interaction with carrier p. in mem
• Rate of diffusion depends on:
• Amount of substrate available
• Velocity of kinetic motion
• No. & sizes of openings thru
which m./ ions can move
• Facilitated: requires interaction of
carrier protein: aids passage of m./
ions thru mem by binding chemically &
shuttling them thru mem
Diffusion of lipid soluble substances: rate:
• Lipid solubility: high: Oxygen, Nitrogen,
Carbon Dioxide, Alcohols
Diffusion of lipid insoluble substances:
water: highly insoluble in mem lipids, but
passes thru channels (eg: aquaporins)
Aquaporins: selectively permit passage of
water thru mem. highly specialized, at least
13 diff types
Water can diffuse rapidly: eg: in RBC per sec
x100 volume of RBC
Small lipid insoluble m. (water soluble) can
pass thru protein pore channels: diameter
increases, penetration decreases rapidly.
Eg: diameter of urea m.: 20% greater than
water, penetration: x1000 less than water
(but due to high water penetration this
amount still allows rapid transport of urea
thru mem)
Selective permeability & Gating of
channels:
Protein pores & Channels: tubular pathways
that span mem
Pores: made of integral mem proteins: form
open tubes thru mem, always open.
Selectivity: due to diameter, shape, nature
of electrical charges & chemical bonds
along inside surfaces
Eg: aquaporins: narrow pore, only permits
water, too small for hydrated ions.
Density of some aquaporins (eg: aquaporin-
2) in cell mem: not static – altered in diff
physiological conditions
Gated channels: regulated by
• Electrical signals: voltage gated
• Chemicals that bind to channel
proteins: ligand-gated
Flexible, dynamic structures, subtle conf.
changes influence gating & selectivity
Selectivity of K+ channels
K+ channels: permit K+ x1000 more readily
than Na+
K+ ions: slightly larger than Na+
X-ray crystallography: K+ channels have
tetrameric structure: 4 identical protein
subunits surrounding central pore.
At top of channel pore: pore loops: form
narrow selectivity filter: lining it: carbonyl
oxygens.
When hydrated K+ enter selectivity filter,
interact with carbonyl oxygens & shed most
of their bound water m. → dehydrated K+
can pass thru channel
Carbonyl oxygens too far to interact with
smaller Na+ ions: effectively excluded by
selectivity filter.

Cation/anion selectivity/ for specific ions:
eg: Na+ channel (d = 0.3-0.5 nm): selectivity
is crucial for proper cell function.
Narrowest part of open pore of Na+ channel
= selectivity filter: lined with strong
negatively charged a.a. residues: pull small
dehydrated Na+ away from hydrating water
m. – these ions don’t need to be fully
dehydrated to pass thru channels.
Gating: means of controlling ion
permeability.
Some gates: thought to be gatelike
extensions of transport protein: can close/
open by conf. change in shape of protein m.
Voltage gating: molecular conf of gate/
chemical bonds responds to electrical
potential across cell mem.
Eg: strong negative charge on inside of cell
mem may cause outside Na+ gates to
remain closed, inside: positive: gates open:
basic mech. for eliciting action potentials in
nerves - responsible for nerve signals.
K+ gates: on intracellular ends, open when
inside of mem is positively charged:
opening of these: to terminate action
potential
Chemical (ligand) gating: binding of
chemical substance → conf./ chemical
bonding change in protein m. → opens/
closes gate
Eg: neurotransmitter acetylcholine opens
gate of acetylcholine receptor: negatively
charged pore: d= 0.65 nm: allows
uncharged m./ small positive ions to pass
thru: important for transmission of nerve
signals: from nerve cell to nerve cell/
muscle to cause contraction.

Shows 2 recordings of electrical current
thru single Na+ channel (~25 mV potential
gradient across mem).
Channel conducts current in an all-or-none
fashion (either open all/almost all of the
time/ closed at diff voltages): at in-btwn
voltages: gates snap open & closed at
irregular intervals → avg current flow btwn
min & max.
Each open state lasts for a fraction of a ms
& up to several ms: rapid changes can occur
Patch Clamp method for recording ion
current flow thru single channels
Micropipette (d = 1-2 μm) placed against
outside of cell mem.
Suction applied inside pipette to pull mem
against top of pipette: creates a seal where
edges of pipette touch cell mem: minute
mem ‘patch’ at tip of pipette thru which
electrical current flow can be recorded.
Small mem patch can be torn away from
cell: pipette with sealed patch is inserted
into a free solution: allows conc. of ions
inside pipette & outside in solution to be
altered as desired.
Voltage btwn 2 sides of mem can be set/
‘clamped’ to a given voltage
Patches can be made small enough so that
only single channel protein is found
By varying conc of diff ions & voltage of
mem: can determine transport
characteristic of the single channel & gating
properties.
Facilitated/ carrier-mediated diffusion:
Carrier facilitates diffusion of substance
• Simple: rate increases proportionately
with rate of diffusing substance
• Facilitated: rate approaches a
maximum: Vmax: as conc increases
Molecule to be transported binds to binding
receptor inside protein carrier (pore)
In a fraction of a sec, conformational/
chemical change occurs in carrier p., pore
now opens into opp side of mem
As binding force of receptor is weak,
thermal motion of attached molecule
causes it to break away & be released on
opp side of mem
Rate can never be greater than the rate at
which the carrier protein molecule can
change back & forth btwn its 2 states.
Note: this mechanism allows molecules to
diffuse in either direction thru mem.
Eg: glucose: 14 mem of mem protein family:
GLUT: transport glucose in various tissues.
Some transport monosaccharides with
similar structures to glucose (galactose,
fructose, etc.)
GLUT 4: activated by insulin, can increase
rate of facilitated diffusion of glucose by
x10-20 in insulin sensitive tissues – principle
mechanism by which insulin controls
glucose use in body
Factors that affect net rate of diffusion:
• Proportional to conc. gradient: rate
of net diffusion into cell:
• Membrane electrical potential &
diffusion of ions – Nernst Potential:
opp. charge attracts, same repels.
When conc diff rises high enough 2
effects balance each other: the
electrical diff that balances given
conc. diff of univalent ions at normal
body temp: Nernst Potential
• Proportional to pressure diff.:
pressure = sum of all forces of diff
molecules striking a unit surface area
at a given instant. (usually higher no.
of molecules = higher pressure):
increased amounts of energy to
cause net movement of m. down
pressure gradient
Osmosis
Volume of cell remains constant as 0 net
movement of water.
Under certain conditions: conc. diff for
water can develop across mem: net
movement occurs → cell swells/ shrinks
Osmosis: net movement of water caused by
a concentration diff of water.
Mem is said to be selectively permeable to
water, less to Na+ & Cl- ions
Osmotic pressure: amount of pressure
needed to stop osmosis
Osmosis from B→ A will cause levels of fluid
columns to become further apart until
pressure diff develops btwn 2 sides of mem
that is great enough to oppose osmotic
effect = osmotic pressure of solution that
contains non diffusible solute
Osmotic pressure exerted by particles
(m./ions) in a solution: determined by no. of
particles per unit volume of fluid (molar
concentration if it is a non-dissociated m,):
each particle exerts on avg same amount of
pressure against mem (larger mass, smaller
velocity: same avg KE)
Osmole: to express conc of a solution in
terms of no.s of particles.
1g molecular weight of osmotically active
solute. (eg: 180g of glucose)
If a solution dissociates into 2 ions, 1g
molecular weight = 2 osmoles (no. of
osmotically active particles = x2)
Solution that has 1 osmole of solute
dissolved in each kg of water: osmolality of
1 osmole/kg
Normal osmolality of extracellular &
intracellular fluids: 300 milliosmoles/ kg of
water
Osmolality & osmotic pressure
At normal body temp, conc of 1 osmole/L
will cause 19,300 mmHg osmotic pressure
in solution (1 milliosmole/L: 19.3 mmHg)
19.3 x 300 (milliosmolar conc. of body
fluids) = total calculated osmotic pressure
of body fluids = 5790mmHg (measured
value = 5500 as many ions in solution are
highly attracted to one another: can’t move
unrestrained/ create full osmotic pressure
potential): actual = 0.93x calculated value
Osmolarity: osmolar conc. expressed as
osmoles per liter of solution rather than
osmoles per kg of water.
Quantitative diff btwn osmolarity &
osmolality < 1% for dilute solutions (in
body) – more practical to measure
osmolarity
Active transport
Movement of ions/ other substances across
mem using carrier protein against an energy
gradient: requires additional source of
energy (besides KE)
Eg: Na+, K+, Ca2+, iron, H+, Cl-, I-, urate ions,
diff sugars, most a.a.
Important to maintain conc. gr. of ions: eg:
energy source must cause excess
movement of K+ inside cells, Na+ to outside
Primary AT & secondary AT
Acc to source of energy used to facilitate
transport
Primary AT: energy derived directly from
breakdown of ATP/ other high-energy
phosphate compound
Secondary AT: derived secondarily from
energy stored in form of ionic conc diff of
secondary molecular/ ionic substances,
created originally by Primary AT.
Carrier proteins in AT functions differently
from carriers in facilitated diffusion (can
move substance against electrochemical gr
using energy)
Sodium potassium pumps: primary AT:
To maintain Na+/ K+ conc gr across cell
mem, establishing negative electrical
voltage inside cells, basis of nerve function
Complex of 2 globular proteins: larger α
subunit (m. weight = 100,000), smaller β
subunit (m. weight = 55,000)
Smaller subunit function unknown: may
anchor protein complex in lipid mem
Larger subunit: 3 specific features:
• 3 binding sites for Na+ on inside
portion of protein
• 2 binding sites for K+ on outside
• Inside portion (near Na+ binding
sites) has ATP activity
Activation of ATPase when 2 K+ & 3Na+
bind → cleavage of one ATP m. → liberates
high-energy phosphate bond: liberated
energy causes conformational & chemical
change in protein carrier: 3Na+ out, 2K+ in
If electrochemical gradients are
experimentally increased to the point that
energy stored in gradients > chemical
energy of ATP hydrolysis, ions will move
down conc. gr, pump will synthesize ATP
(phosphorylated form of pump donates
phosphate to ADP → ATP)
Relative conc. of ATP, ADP, phosphate,
electrochemical gradients of Na+ & K+
determine direction of enzyme reaction
For electrically active nerve cells: 60-70%
cell’s energy requirement: for pumping Na+
out, K+ in
Functions: Control cell volume (otherwise
cells will swell & burst): inside cell proteins,
organic molecules that can’t escape: usually
negatively charged, so attract K+, Na+,
other positive ions: cause osmosis of water
in – Na+-K+ pump: 3 ions out for 2 in (net
loss out → osmosis out) & as Na
permeability is lower than K+ so strong
tendency to remain outside cell
Calcium pump: primary AT:
Ca2+ normally maintained at v. low conc in
intracellular cytosol (x10,000 less than ECF)
2 primary AT Ca2+ pumps:
• In mem: pumps Ca2+ outside cell
• Pumps Ca2+ into one/more
intracellular vesicles (eg: SR in m.,
mitochondria in all cells)
Carrier protein penetrates mem, functions
as enzyme ATPase, specific binding site for
Ca2+
Primary AT of H+
Important at:
• Gastric glands in stomach (deep lying
parietal cells): for secreting HCl: at
secretory ends [H+] increased to a
million-fold → then released into
stomach with Cl- to form HCl
• Late distal tubules & cortical collecting
ducts of kidneys: special intercalated
cells: large amounts of H+ secreted from
blood into renal tubular fluid (against
conc. gr. of x900) to eliminate excess H+
from body fluids
Energetics of primary AT
Amount of energy to transport substance
actively thru mem determined by substance
conc.
Energy required: proportional to log of the
degree of conc.
Eg: to concentrate 1 osmole of a substance
10-fold is about 1400 calories, 100-fold:
1400 x2 = 2800 calories
Some cells (eg: cells lining renal tubules &
many glandular cells): expend ~90% energy
for moving substances against conc. gr.
Secondary AT: Co-transport & Counter-
transport
Sodium gradient = storehouse of energy
(excess Na always tries to diffuse in): can
pull other substances with Na thru cell mem
= co-transport
Coupling mechanism required: achieved by
another carrier protein: attachment point
(has 2 external binding sites) for both Na+
and transported substance
Eg:
• Sodium-glucose transporters: across
renal & intestinal epithelial cells
• Sodium co-transport of aa: 5 aa.
transport proteins identified: each
transports one subset of a.a with
specific molecular characteristics
• Co-transport of K+, Cl-, bicarbonate,
phosphate, iodine, iron, urate
Counter-transport: Na+ binds to carrier
where it projects to exterior surface of mem
& transported substance binds to interior
projection → conf change → energy
released by action of Na+ moving in used to
move substance out
Eg:
• Sodium-calcium counter-transport:
thru almost all mem
• Sodium-hydrogen counter-transport:
in proximal tubules of kidneys: can
transport extremely large numbers of
H+: key to H+ control in body fluids
Note: for both co- & counter- transport
conf. change occurs after BOTH m. have
bound
Active transport thru cellular sheets
• Intestinal epithelium
• Epithelium of renal tubules
• Epithelium of all exocrine glands
• Epithelium of gall bladder
• Mem of choroid plexus of brain
Basic mechanism: AT thru cell mem on one
side of transporting cells in sheet, then
simple/ facilitated diffusion thru mem on
opp. side of cell
Eg: Na+ thru epithelial sheet of intestines/
gall bladder/ renal tubules:
Epithelial cells connected together tightly at
luminal pole by means of junctions.
Brush border on luminal surfaces:
permeable to Na+ & water both: diffuse
from lumen to interior of cell
At basal & lateral mem: Na+ actively
transported into extracellular fluid of
surrounding CT & blood vessels:
Creates high Na+ ion conc. gr. across mem
→ causes osmosis of water
Thru these mechanisms: almost all
nutrients, ions, other substances absorbed
into blood from intestine, reabsorbed from
glomerular filtrate by renal tubules