the structure of the cell membrane resting membrane...
TRANSCRIPT
04/10/2013
1
The Structure of the Cell
Membrane
Resting Membrane Potential
10.09.2013.
� Structure of the cell membrane.
� Resting membrane potential.
• The Nernst equation.
• Donnan potential.
• The Goldman-Hodgkin-Katz equation
Phospholipids
Polar – head(hydrophilic)
Non-polar – tail
(hydrophobic)
⇒⇒⇒⇒ „water soluble fat”
phosphatidil – cholin0
The main component of the biological membranes.
Phospholipid = diglyceride (glycerine+fatty acid) + phosphate group + organic molecule (e.g. choline).
04/10/2013
2
Irving LangmuirAmerican physico-chemist
1932 Nobel-price in chemistry
• 1917 – lipids form a monolayer on thesurface of the water� polar heads (hydrophilic) – oriented
toward the water� nonpolar tails (hydrophobic) – oriented
away from waterwater
Irving Langmuir, "The Constitution and Fundamental Properties of Solids and Liquids. II," Journal of the American Chemical Society 39 (1917): 1848-1906.
Lipid bilayer
1925 – Evert Gorter & F. Grendel (University of Leiden, Holland)
• Compared the measured surface area of the erythrocytes and the surface area
calculated from the lipid content of them.
• Gorter E, Grendel F. On Bimolecular Layers of Lipoids on the Chromocytes of the
Blood. J Exp Med. 1925 Mar 31;41(4):439-43.
Gortel, E. & Grendel, F. (1925) On bimolecular layers of lipoid on the chromocytes of the blood. J. Exp. Med. 41, 439–443.
04/10/2013
3
Lipid bilayer
• twice as much lipid in the membrane of the red blood cells than needed for a monolayer → lipid bilayer
IC
EC
Polar heads toward the intra- and extracellular
space
Apolar tails in the middle
Gortel, E. & Grendel, F. (1925) On bimolecular layers of lipoid on the chromocytes of the blood. J. Exp. Med. 41, 439–443.
04/10/2013
4
Gibbs free energy [Joule]
G = H - TSA spontaneous process is accompanied bya decrease in the Gibbs energy at constanttemperature and pressure.
At constant temperature and pressure thechange in the Gibbs energy is equal to themaximum non-expansion workaccompanying a process.
Hydrophobic interaction
• hydrophobic = water-repelling; low affinity (solubility) for water
• Walter Kauzmann (American chemist) - Nonpolar molecules in polar
environment (solvents) are trying to minimize their contact with water
• 1”cage” formation → 2clustering
• Factors affecting the strength of hydrophobic interaction
– Temperature (T ↑ ⇒ Strength ↑)
– Number of carbons in the hydrophobic molecule (Length ↑ ⇒ Strength ↑)
– Number of “non single” bonds (e.g. double, triple bonds…) in the hydrophobic molecule
(shape) ( # “non single” bonds ↑ ⇒ Strength ↓)
04/10/2013
5
Thermodynamic changes
hydrophobic
molecule
H2O
+ hydrophobic
molecule
H2O
→
H2O H2O
hydrophobic
molecule
hydrophobic
molecule
Clustering
(forming hydrophobic interactions)
∆H = small positive
∆S = large positive
∆∆∆∆G = negativeSPONTANEOUS PROCESS
Cage formation
(no interaction between hydrophobic molecules)
∆H = small positive
∆S = large negative
∆∆∆∆G = positiveNON SPONTANEOUS PROCESS
„Fluid mosaic” model
• phospho-lipid bilayer
• Fluid – lateral movement of the components („floating”)
• Mosaic – the mosaic-like arrangement of themacromolecules
http://www.molecularexpressions.com/cells/plasmamembrane/plasmamembrane.html
• 1972 - Singer and Nicholson „fluid mosaic” model
Singer SJ, Nicolson GL. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(23):720-31.
04/10/2013
6
Structure of the cell membrane
rotation
Polar
(hydrophilic) head
Non-polar
(hydrophobic) tail
Lateral diffusion
Flip-flop~ 5 nm
Protein molecule (~30-50%)
Phospholipide molecule (~40-60%)
Functions of the membrane poteins
• Ion channels (Na+/K+ ATPase; K+ channel…)
• Transporters (Aquaporin-H2O transport)
• Structural elements
• Intracellular connections (anchoring – cytoskeleton)
• Extracellular connection (gap junction: cell to cellcontact between cardiac cell)
• Signal transduction (action potential)
• Receptors (insulin receptor)
04/10/2013
7
The main components of the intra- and extracellular space
• water
• Ions
– Kations (K+, Na+, Ca2+)
– Anions (Cl-, H2PO4− and HPO4
2− ions)
• proteins
– Mainly intracellular localisation
– Negatively charged polyvalent (having more than one
valence) macromolecules (pH! – isoelectric point)
Membrane potential
The electrical potential difference (voltage)
across a cell's plasma membrane.
Extracellular space
Intracellular space
0V
Microelectrode
-100 mV > Uresting < -30 mV
04/10/2013
8
Ionic concentrations inside and outside of a muscle cell
Na+ : 120 mMK+ : 2.5 mM
Cl- : 120 mM
Na+ : 20 mM
K+ : 139 mM
Cl- : 3.8 mM
Forces controlling the movements of charged particles
Chemical potential energy:
� The chemical potential of a thermodynamic system is the amount of energy
(Joule) by which the system would change if an additional particle were
introduced (~ number of the particles!).
� Concentration gradient → diffusion: moving the particles through the
permeable membrane from a high concentration area to a low
concentration area → diffusion potential.
Energy: Capacity for doing work.
04/10/2013
9
Electric potential energy
• the result of conservative Coulomb forces
• associated with the configuration of a particular
set of point charges within a defined system
• work required by an electric field to move
electric charges (Joule).
• Electrical gradients: The sum of the “+” and “-”
are not the same at the different points in space.
• An electric field creates a force that can move
the charged particles (the work of the electric
field) → moving charged particles = electric
current.
K+ : 100 mM
Cl- : 100 mM
K+ : 5 mM
Cl- : 5 mM
Force controlling the movements of ions through the cell membrane
Electro-chemical potential
= the combination (sum) of the chemical and the electric
potential energy.
04/10/2013
10
Julius Bernstein (1839 - 1917) - German physiologist
1./ The cell membrane is selectively permeable to potassium• Ca2+ sensitive potassium channels• Inwardly rectifying potassium channels• Voltage-gated potassium channels• “Tandem pore domain potassium channel” – “leak channel” (K2p)
1952: Hodgkin and Huxley suggested the leakage of current etchum, KA; Joiner, WJ; Sellers, AJ; Kaczmarek, LK; Goldstein, SA. (1995) A
new family of outwardly rectifying potassium channel proteins with two poredomains in tandem. Nature, 376 (6542): 690-5.
2./ The intracellular potassium cc. is high
3./ The extracellular potassium cc. is low
Bernstein’s potassium
hypothesis (1902)
Bernstein,J.(1902).Untersuchungen zur Thermodynamik der bioelektrischen Strome. Pflugers Arch.ges. Physiol. 92, 521–562.
Bernstein’s potassium hypothesis
K+ : 100 mM
Cl- : 100 mM
K+ : 5 mM
Cl- : 5 mM
04/10/2013
11
Bernstein’s potassium hypothesis
K+ gradient (chemical potential)
electric gradient
(electrical potential)
The side with
high
concentration of
positive ions
becomes the
negative side !!!!
[K+] [K+]
[Cl-] [Cl-]
- +
How is it possible to quantify the Bernstein’s hypothesis ?
(calculating the electrical potencial (value, number)
04/10/2013
12
Walther Hermann NernstGerman physical chemist
(June 25, 1864 – November 18, 1941)
Calculating the electrical potential at which
there is no longer a net flux (movement) of a
specific ion across a membrane.
N = number of moles associated with the concentration gradient
R = gas constant
T = absolute temperature
X1 / X2 = concentration gradient
N = number of moles of the charged particles
z = valency (number of + or – charges (e.g. K+ : monovalent))
F = Faraday’s number
E = strength of the electric field = electric potential or electrostatic potential
= The work needed to move a unit electric charge from one point to another
against an electric field (Joule/Coulomb = Volt).
Electric potential energy ⇒ Welectr=NZFE
Chemical potential energy ⇒ Wchem=NRTln��
��
04/10/2013
13
Equlibrium (resting) condition
2
1ln
X
XNRTNzFE =
2
1ln
X
XRTzFE =
2
1lnX
X
zF
RTE =
Electrical potential energy Chemical potential energy
Equlibrium potential
Nernst equation: What membrane potential
(E) can compensate (balance) the
concentration gradient (X1/X2).
2
1lnX
X
zF
RTE =
The inward and outward flows of the ions are balanced
(net current = zero → equilibrium = stable, balanced, or unchanging
system).
04/10/2013
14
Nernst equation
( )( )out
inmV
C
C
zE log
58−=
2
1ln
X
X
zF
RTE =
Ionic concentrations inside and outside of a muscle cell
[K+] ⇒ EmV = -58/1 log (139/2.5) = - 101.2 mV
[Na+] ⇒ EmV = -58/1 log (20/120) = + 45.1 mV
[Cl-] ⇒ EmV = -58/1 log (3.8/120) = + 86.9 mV
Na+ : 120 mMK+ : 2.5 mM
Cl- : 120 mM
Na+ : 20 mM
K+ : 139 mM
Cl- : 3.8 mM
EmV=-92mV= 30.8 mV
04/10/2013
15
What happens if the cell
membrane is permeable to
more than one type of ion?
Frederick George Donnan
Donnan equilibrium: characterising the equlibrium
situation when the membrane is not permeable for
some ionic components.
- non-moving charged component (e.g. intracellular
proteins) → equlibrium concentration difference
- more than one diffusible ion (K+, Cl-)
(1870-1956; Irish chemist)
04/10/2013
16
Donan equlibrium - at start
[K+]
[Pr -] [Cl-]
[K+]
K+=x K+=x
Pr-=x Cl-=x
Cl- concentration gradient
K+ concentration gradient
Cl- electrical gradient
K+ electrical gradient
A B
A B
Donan equlibrium - at equlibrium
[K+]
[Pr -] [Cl-]
[K+]
K+=x+y K+=x-y
Cl-=y Cl-=x-y
Pr-=x
A (-) B (+)
[Cl-]
A B
- +
04/10/2013
17
Donnan rule of equilibrium
• Diffusible ions: K+, Cl-
• In equlibrium the elektro-chemical potentials are equal.
[ ][ ]
[ ][ ]
in
out
out
in
Cl
Cl
zF
RTE
K
K
zF
RTlnln ==
[ ][ ]
[ ][ ]
in
out
out
in
Cl
Cl
K
K=
[ ][ ] [ ][ ]outoutinin
ClKClK =
The Donnan rule is valid only when the ions are passively distributed.!
The Gibbs–Donnan equilibrium is a phenomenon that contributes to the formation of
an electrical potential across a cell membrane.
What happens if the Donnan rule is not obeyed?
04/10/2013
18
Goldman-Hodgkin-Katz Constant
field equation (Goldman equation)
To determine the potential across a cell's membrane taking intoaccount all of the ions with different permeabilities through themembrane.
David E. Goldman (USA)
Alan Lloyd Hodgkin (England)
Bernard Katz (England).
The Goldman equation for M positive ionic species and A negative:
•Em = The membrane potential•Pion = the permeability for that ion•[ion]out = the extracellular concentration of that ion•[ion]in = the intracellular concentration of that ion•R = The ideal gas constant•T = The temperature in kelvins•F = Faraday's constant
Goldman equation
A "Nernst-like" equation with terms for each permeant ion.- All the ions are involved.- Good agreement with the measured values (muscle cell: Umeasured=-92mV_Ucalc.=-89.2mV).
[ ] [ ]
[ ] [ ]
+
+=
∑∑
∑∑−+
−+
−+
−+
M
j outjA
N
i iniM
M
j injA
N
i outiM
m
APMP
APMP
F
RTE
ji
ji
ln
04/10/2013
19
The membrane potential is the result of a„compromise” between the various equlibriumpotentials, each weighted by the membranepermeability and absolute concentration of theions.
Goldman equation
The end!