Dr. Assaad. A.

Eid
[email protected]
 Cell Membrane

Ions Cannot Diffuse Across the Hydrophobic Barrier of the Lipid Bilayer
 Ion Channels

q Ions Cannot Diffuse Across the Hydrophobic Barrier of the Lipid Bilayer
q Ion Channels Provide a Polar Environment for Diffusion of Ions Across
the Membrane
 Ion Channels

–
² membrane proteins found in all cells of the body

² structurally quite heterogeneous so that different types of

channels in different part of the system can carry out specific
signaling tasks

² Malfunctioning causes diseases: cystic fibrosis; cardiac

arrhythmia, etc.

² can be site of action of drugs, poisons and toxins

Specialized Functions of Ion Channels

–
q Mediate the generation, conduction and transmission of electrical
signals in the nervous system
q Control the release of neurotransmitters and hormones
q Initiate muscle contraction
q Transfer small molecules between cells (gap junctions)
q Mediate fluid transport in secretory cells
q Control motility of growing and migrating cells
q Provide selective permeability properties important for various
intracellular organelles
 Properties of Ion Channels:
–
Three Important Properties of Ion Channels:

q Selective, they recognize specific ions or chemicals;

q Conduct ions
v channels conduct so fast (108 ions/sec)
v channels act as catalyst

q Gating
v They open and close in response to specific electrical,
mechanical or chemical signals:
 Non-Gated Ion Channels

–
² NON-GATED CHANNELS

q Plasma membrane has many non-

gated K+ channels (some of them
‘leaky’) allowing a slow facilitated
diffusion of K+ out of the cell

q Permeability K+ >> Na+

 Gated Ion Channels

–
² GATED CHANNELS

q open or close in response to stimuli

q stimulated by:
v Ligand binding (chemicals) - Ligand-gated (nAChR, GABAAR,
GluR, glyR – many subtypes within each category)

v Voltage (Membrane potential) - Voltage-gated (Na+, K+ - many

subtypes within each category)

v Mechanical force (eg. membrane stretch)

v Phosphorylation
Ligand Gated

–
Extracellular
Voltage Gated Channels

–
Stretch Gated Channels

–
Phosphorylation

–
Modifiers of Channel Gating
–
 Thick Ascending Limb (TAL)

–
LUMEN BLOOD
Na+
K+
Na+
2Cl- +
K
Cl-
HCO3-
K+ K+

Cl-

At least 25% of filtrate is reabsorbed via Na+-2Cl--K+ co-transporter in luminal membrane of this segment.
 Distal Convoluted Tubule (DCT)

–
LUMEN

Na+
Na+ K+
Cl-

K+

Cl-

BLOOD
Up to 5%-8% of filtrate is reabsorbed actively via NaCl co-transporter in apical cell membrane.
 Major Characteristics of Ion
Channels
–
q The Flux of ions through the ion channel is passive.

v does not require energy

v depends on electrical and chemical forces across
membrane

q Channels select the type of ion that is allowed to cross the

membrane

v most cationic channels are primarily selective to

one cation
v most anionic channels are highly selective
 Selectivity

–
q Highly selective: based on specific interaction (Na+, K+, Ca2+,
Cl-)

q Moderately Selective: charge based (cation or anion)

q Low selectivity: size specific: based on the subunits forming

the channels (gap junctions)
Ion channels vs. Ion transporters

–
 TWO FACTORS DETERMINE
FLOW OF IONS IN HIGHLY SELECTIVE CHANNELS

–
q Size of the ion (With Its H2O Shell) Relative to Channel Pore
Size

q Chemical Interaction of Hydrated Ions With Charged

Amino Acids in Channel walls
 SIZE OF THE ION CHANNELS

–
q Size of the ion (With Its H2O Shell) Relative to Channel Pore
Size
v to explain why some K+ channels allow K+ ions to pass through
and not Na+:
Knowing that K+ is larger than Na+, but Na+ ion has
more highly localized charge and, therefore, stronger
electric field; it thus attracts H2O more strongly and
therefore has larger H2O shell
v an ion with smaller H2O shell moves faster in solution

This will explain why some channels let the K+ go through but
not the Na+
CHEMICAL INTERACTION WITH CHANNEL WALLS

–
How to explain why some channels allow Na+ to pass through and not K+
ions

q Chemical Interaction of Hydrated Ions With Charged Amino

Acids in Channel walls
 Ion Selectivity: Chemical Interaction of Hydrated Ions
With Charged Amino Acids in Channel walls

–
q Channels are made up of proteins they
have narrow regions (molecular sieves)
act as selectivity filters with polar
(charged) amino acids that line the filter.

q At this site an ion shed most of its water

of hydration and, in their place, form
week chemical bounds with polar amino
acid residues that line the walls of the
channel. Ion will traverse a channel only
if the energy of interaction will
compensate for the loss of waters of
hydration.
 Ion Selectivity: Chemical Interaction of Hydrated Ions
With Charged Amino Acids in Channel walls

–
q The selectivity is dictated by the negative
field strength of the ion wall:
v High negative field strength (formed
by negatively charged carboxylic acid
groups of glutamate or aspartate) will
selectively binds to Na+.
v low field strength (formed by polar
hydroxyl oxygen or carbonyl atoms
would select K+.

q Ions bind to the selective filter for very

short time, after which the electrostatic
and diffusional forces propel the ion
through the channel
 CHANNEL GATING
(Opening & Closing of Channels)

–
q The channel protein has two or more conformational states that are
relatively stable.

q Ion channels have at least one open state and one or two closed states
v resting (or activable/closed)
v active (or open)
v refractory (or inactivable)

q The transition of the channel between these different states is called

gating and it involve a temporal change in the molecular structure.
 Factors Involved in Gating

–
q Rapidity of gating dependent on the metabolic state of the
cell

q Energy need to be supplied for changing from closed to

open states (electrical, chemical or mechanical energy
dependent on the regulatory mechanism)

q Signals that gate channels also control the rate of

transition between open and closed states
Models of Gating

–
 Voltage Gated Channel

–
Energy provided by the movement of a charged region of the
channel protein called “voltage-sensor” through the membrane
electric field. It can sense the changes in membrane voltage
v act on a charged component of the channel
 Voltage-Gated Ion Channels

q Open and close in response to voltage change

q Found in cardiac, skeletal, and neuronal cells
q Voltage change causes membrane potential difference that can lead
to depolarization
q Can produce an action potential
q Share the same basic structural themes.
 Ligand Gated Channel

channel ligands could be:

–
v extracellular (eg. neurotransmitters, hormones) or
v intracellular include:
1. cytoplasmic constituents (eg. Ca+2, nucleotides)
2. activation of a cascade leading to covalent modification
of channel by phosphorylation
 Stretch Gated Channel

–
Energy supplied by mechanical stretch or pressure
resulting in
v mechanical forces on the cytoskeleton or
v a change in the tension of the adjoining lipid bilayer
 STRUCTURE OF CHANNELS

–
q In addition to the biophysical properties described
from the patch clamp technique

q All channels consist of a large glycoprotein (a

protein with a CHO attached)

v made of two or more primary

subunits, either identical or different
v each subunit spanning the entire
membrane width
v with the subunits cooperating to form
the aqueous channel pore

q Some channels have auxiliary subunits

(cytoplasmic or membrane embedded)
 Patch Clamp

–
q With this method, the ionic current is measured on a tiny
membrane patch to which a pre-determined voltage-clamp is
applied

q It can be used to measure currents passing through individual

ion channels or through whole cells
 Patch Clamp Measurements

–
Cell-attached (on-cell): the pipette
makes a tight contact with the intact
cell allowing measurements of single-
channel currents

Inside out: upon cell-attached

configuration the pipette is withdrawn
while the contact is maintained. The
inside (cytoplasmic) side of the
membrane is facing the bath fluid. This
configuration is used for single-
channel recordings
 Patch Clamp

–
The patch clamp determines:

q the molecular composition of ion channels

q when a single ion channel opens or closes

q the changes in configuration upon opening and closing

q channels specificity towards certain ions

Transporters
 Overcoming the Cell Barrier

– Cell
q The cell membrane is a barrier, but:

v Nutrients must get in

v Products and wastes must get out

q Permeability determines what moves in and out of a cell

q A membrane is:

v Impermeable if it lets nothing in or out

v Freely permeable if it lets anything pass

v Selectively permeable if it restricts movement

 Concept of Biological Barrier

–
q Biological barriers arise because of the lipid bilayer nature of the cell
membrane (which allows only lipophilic substances or non polar to diffuse
through the lipid phase).

q Cell membranes are selectively permeable:

v Allow some materials to move freely

v Restrict other materials

q The selective permeability properties is a function of the presence of

specialized protein transporters embedded into the membrane (channels,
carriers, or pumps) that allow the transport of specific hydrophilic solutes.
 Diffusion in Solutions

q All molecules are constantly in motion

–
v Molecules in solution move randomly

v Random motion causes mixing

q Concentration is the amount of solute in a

solvent

q Concentration gradient:

v More solute in one part of a solvent than

another

q Solutes move down a concentration gradient:

v Molecules mix randomly

v Solute spreads through solvent

v Eliminates concentration gradient

 Factors Affecting Diffusion Rates

–
q Distance the particle has to move

q Molecule size:

v Smaller is faster

q Temperature:

v More heat, faster motion

q Gradient size:

v The difference between high and low concentration

q Electrical forces:

v Opposites attract, like charges repel

 Diffusion vs. Osmosis
–

q Diffusion: movement of solutes across cell membrane

q Osmosis is the movement of water across the cell membrane
 KEY CONCEPT

–
q Concentration gradients tend to even out

q In the absence of a membrane, diffusion eliminates concentration

gradients

q When different solute concentrations exist on either side of a

selectively permeable membrane, osmosis moves water through the
membrane to equalize the concentration gradients
 Transport Through Cell Membranes

–
q Transport through a cell membrane can be:

v Active (requiring energy and ATP)

v Passive (no energy required)

q 3 categories of transport

v Diffusion (passive)

v Carrier-mediated transport (passive or active)

v Vesicular transport (active)

 Passive Transport: Diffusion and the Cell
Membrane
–
q Diffusion can be simple, channel, or carrier mediated
q Carrier mediated diffusion is:

v Specific

v Subject to saturation
 Mediated-Transport Systems

–
q Many molecules (like glucose) are either too large and charged to
get into the cell without help.

q The protein transporters (also called carriers) bring these

molecules into and out of cells by conformation changes.

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 Mediated Transport System

In both simple and

facilitated diffusion,
solutes move in the
direction predicted
by the electrochemical
gradient.
 Mediated Transport System

In both simple and facilitated

diffusion, solutes move in the
direction predicted by the
electrochemical
gradient.

In active transport, solutes

move opposite to the
direction predicted by the
electrochemical gradient.
 Transporters

–
q Transporters are specific for their ligand.

q Transporters do not move as many molecules as channels

do because of binding and conformational shifts.

q Transporters can be saturated. This means that there is a

maximum flux of molecules that can be reached.
 Active Transport

–
q Active transport uses energy to move molecules against
the concentration gradient.

q These transporters are often called “pumps”.

q These pumps can also be saturated and use two types of

energy sources:

v The direct use of ATP in primary active transport

v The use of an electrochemical gradient across a
membrane to drive the process in secondary active
transport
 Sodium-Potassium Exchange Pump

–
q Active transport, carrier mediated:
q 1 ATP moves 3 Na+ out 2 K+ in
q This creates an electrical potential across the membrane
v Called the Transmembrane Potential
 Primary Active-Transporters

–
q The Na+/K+-ATPase primary active transporter is found in
every cell and helps establish and maintain the membrane
potential of the cell.

q In addition to the Na+/K+-ATPase transporter, the major

primary active-transport proteins found in most cells are:
v Ca2+-ATPase
v H+-ATPase
v H+/K+-ATPase
 Secondary Active Transport

–
q Secondary active transport is
distinguished from primary
active transport by its use of
an electrochemical gradient
across a plasma membrane
as its energy source.

q Transporters that mediate

secondary active transport
have two binding sites, one
for an ion (e.g., Na+)and
another for the co-transported
molecule (e.g., Glucose).
Secondary Active Transport

–
 Secondary Active Transport

q Cotransporters (symporters) move molecules in the same direction.

q Countertransporters (antiporters) move molecules in opposite directions.

Membrane Transport Proteins

–
 Vesicular Transport

q Also called bulk transport

–
q Transport of large particles and macromolecules across
plasma membranes
Directional Descriptive Terms
v Exocytosis – moves substance from the cell interior to
the extracellular space
v Endocytosis – enables large particles and
macromolecules to enter the cell
™ Receptor-mediated
™ Pinocytosis(fluid)
™ Phagocytosis
Functional Descriptive Terms
v Transcytosis – moving substances into, across, and
then out of a cell
v Vesicular trafficking – moving substances from one
area in the cell to another
v Phagocytosis – pseudopods engulf solids and bring
them into the cell’s interior
 Endocytosis

–
Ø There are three general types of endocytosis that may occur in a cell:
1. Fluid endocytosis (pinocytosis)
2. Phagocytosis
3. Receptor-mediated endocytosis

56
 Exocytosis

–
Movement of molecules out of the cell via vessicles.

57
 Exocytosis

–
Exocytosis performs several functions for cells:
1. Provides a way to replace portions of the plasma
membrane that endocytosis has removed
2. Adds new membrane components to the membrane
3. Provides a route by which membrane-impermeable
molecules (such as protein hormones) the cell synthesizes
can be secreted into the extracellular fluid

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