15.

TRANSPORTERS & ION CHANNELS I

1. TRANSPORT ACRCOSS AN IMPERMEABLE BARRIER
• Transport of ions and small molecules is important for homeostasis and
function of the cell
• Multi-pass transmembrane proteins serve as transporters and ion channels
• Create hydrophilic microenvironments
• Allow continuous of pathway across the plasma membrane for solutes
(hydrophilic)

3 General Principles
• Relative permeability of molecules (gases, water,
glucose)
• Concentration Gradient vs. Electrochemical
Gradient
• Gradients store potential energy

2. ION CONCENTRATIONS
• How are the concentrations of different ions
maintained?
• Store potential energy
By active transport pumps
3. MECHANISM OF TRANSPORT
1. Simple Diffusion: Direct movement of solutes across the membrane
gases, water (by osmosis)

2. Passive Diffusion: Movement driven by gradient (concentration gradient,

potential energy, electrochemical gradient) and facilitated by membrane
proteins
ions, water

3. Active Transport: Movement facilitated by membrane proteins and requires

ATP or Ion Gradient (Primary or Secondary)
Transport is directional, since it coupled to a metabolic energy source
ions, glucose

Passive Transport: Carrier Mediated Transport (Downhill)

• All transporters allow solutes to cross the membrane only passively
• Generally, carrier mediated transport has one or more specific binding sites
• Change between 2 conformational states, and the process is reversible
o Outward-opening à Occluded à Inward Opening
• Net movement of transported solutes depends on the
concentration/electrochemical gradient
o Remember gradient drives movement*
Active Transport (Uphill)
1. Primary Active Transport: Transport facilitated by ATP-driven pumps, where
energy is derived from ATP hydrolysis

2. Secondary Active Transport: Coupled transporters (symport or antiport);

Harness energy stored in a gradient to couple the uphill transport of one
solute across the membrane to the downhill transport of another. Indirectly
uses ATP. Uses the free energy released during the movement of an
inorganic molecule down its concentration gradient as a driving force to pump
the other solute uphill

Symporters (co-transporters) couple the uphill transport of a solute to the

downhill of a solute moving in the same direction
Antiporters (exchangers) couple the uphill transport of a solute to the downhill
transport of a solute that is moving the opposite direction

EXAMPLE
Na+ - GLUCOSE SYMPORT
• Many symporters are driven by the concentration gradient of Na+
o Tends to move into the cell, down its concentration gradient
• Binding of Na+ in State A, promotes binding of glucose
• Binding of both Na+ and Glucose causes a conformational change to State B
of membrane protein
• Both are then released into cytosol
• Na+ gradient drives transport of glucose (Na+ is moving towards it’s
concentration gradient, glucose being dragged against its own)
o Binding of the two solutes is cooperative à the binding of one solutes
increase the protein’s affinity to
the other
4. H+ Regulate Intracellular pH
• Ion gradients (Na+) can be used to transport H+
• Theses exchangers are electroneutral, yet pH is regulated
•

TRANSPORT ATPases
5. P-TYPE ATPase
Na+-K+ ATPase
• P(phosphorylation)-type ATPase
• Function dependent on phosphorylation of intracellular aspartic acid residue,
and ATP hydrolysis
• Can use up 2/3 of cell’s ATP
• These types of transporters are important for setting up and maintaining
gradients of Na+, K+, H+ and Ca2+

Ca2+ ATPase
• Cytosolic [Ca2+] must be kept low
o This related to calcium signaling, where a steep gradient is set up so
when an extracellular signal is present, flow Ca2+ down its steep
concentration allows for rapid signal transmission
• 2 mechanisms of how this is accomplished is by a pump in the PM and ER
SR Ca2+ ATPase
• Contains 10 alpha-helices
o 3 are to line a “channel” that spans the membrane
o 2 are to interact directly with Ca2+
• Ca2+ binding triggers a conformational change that closes the passage way to
the cytosol and activate a phosphotransfer-reaction from ATP (bound to a
nucleotide-binding domain) to an aspartic residue
• ADP is then replaced by an ATP which induces a conformational change that
allows passage to the SR lumen
• 2 Ca2+ ions exits
• Entry of 2 H+ induces a conformational change to close entry into lumen
•
6. V-TYPE ATPase
• Turbine-like protein machine, constructed from multiple different subunits
• Uses ATP molecule as energy source to transport H+ into organelles
(lysosomes, vesicles) to acidify the interior of the organelle

7. F-TYPE ATPase (ATP Synthases)

F0F1-ATPase
• Uses proton gradient as energy source to produce ATP molecule
o Works in reverse from V-Type
• Structurally similar to V-Type ATPases

8. ATP-BINDING CASETTES (ABC) TRANSPORTERS

• 2 domains of 6 transmembrane helices, allow form the structure through
which the molecule passes through the membrane
• 2 intracellular domains for ATP binding
o ATP binding induces dimerization of these ATP binding domains
o Hydrolysis induces release
• Dimerization induces a conformational change that allows the complex for
transport
• Export amino acids, sugars and inorganic ions, proteins
9. AQUAPORINS
• Initially discovered in RBC
• Important for regulating cell size and water content
• Transmembrane proteins with several hydrophilic alpha-helices
• 4 identical subunits, with a central channel
• Very selective (prevent even H3O+ from passing through)
o Normally H+ ions diffuse through water very rapidly by molecular relay
mechanism, by making and breaking hydrogen bonds between each
adjacent water molecule. (Recall from Organic Chemistry)
o 2 central Asparagine are strategically placed to occupy the valence
electrons of the water molecules, therefore preventing the central water
molecule from participating in the H+ relay.
10. SUMMARY*
Classes of
ATPase Solute Location Function

Pumps Na+ out

P-type Plasma and K+ into
ATPase Na+ and K+ Membrane cytosol
Pumps Ca2+
Ca2+ Plasma out of the
Membrane cytosol
Sarcoplasmic Pumps Ca2+
Ca2+ Reticulum into the
cytosol
Pumps H+ into
H+ Membranes of the
organelle and organelle to
V-type secretory acidify the
vesicles environment
Proton
F-type H+ Mitochondrial gradient
ATPase Membrane IM drives ATP
synthesis
Many ions,
sugars,
ABC amino acids, Plasma
Transporters proteins Membrane