Bio>sico-Qumica

2016
Transporte a travs de
membranas

Ernesto Ambroggio (ernesto@mail.fcq.unc.edu.ar)

Qu, cmo y porqu?

Bio>sico-qumica del ujo de molculas a
travs de la membrana celular
Qu se transporta?

Cmo atraviesa la Mb

Difusin simple

Difusin simple,
difusin facilitada,
transporte activo

96

Transporte Pasivo

CHAPTER 4

Cell Membrane Transport

__

0
j

j = + RT lnC j + Z j F +V j P + m j gh

Chemical driving
force

(a)

causes this unequal charge distribution until Chapter 7, when

we explore the origin of membrane potentials in neurons.) Once
we have a firm understanding of what a membrane potential is, we
Las molculas se mueven a travs de la
then consider its influence on the forces that affect ion transport.

membrana como consecuencia de su

The Membrane Potential The fluids in the body contain
gradiente de potenciales qumicos o
a wide variety of solutes, including many ions (also called
electroqumicos.
electrolytes)that is, substances possessing an electrical charge.
Some ions are cations, which have a positive charge; others are
Es el resultado de la produccin de entropa
anions, which have a negative charge. Ions are also present in salt sodonde, al nal, hay ms desorden (cese de
lutions, such as seawater, but we normally cannot detect the presence
ujo) of the ions electrical charges because the number of positive charges

matches the number of negative charges. Such a solution is said to be

electrically neutral because the positive and negative charges cancel
out each other, giving a net (total) electrical charge of zero. Likewise,
the total electrical charge of your body is zero because the number
of cations in your body equals the number of anions. A person may
i when2you rub your
1 feet
pick up and emit charges, however, such as
on a carpet (pick up negative charge) and then touch a metal object
or other conducting material to release this electrical charge.
In intracellular or extracellular fluid, cations and anions are
i i
present in unequal numbers; consequently,
these fluids are not
i
electrically neutral. Intracellular fluid contains a slight excess of
anions over cations, giving it a net negative charge. Extracellular
fluid contains a slight excess of cations over anions, giving it a net
positive charge. Because positive and negative charges are distributed unequally between the inside and outside of a cell, a separation of charge is said to exist across the membrane ( Figure 4.2 ).

J = P (C C

Chemical driving
force

(b)

Figure 4.1 Chemical driving forces. (a) When the concentration of

KD
P=
d

Excess
anions

Excess
cations

98

Transporte Pasivo-Iones
CHAPTER 4

Cell Membrane Transport

Electrical driving force

Vm = 50 mV
(a)

zF
J = P C
C
RT

larger equilibrium potentials because a greater electrical force is

required to equal or balance a larger chemical force. The sign of
the equilibrium potential is such that the electrical force goes in
the directioni opposite to the chemical force. In the case of sodium
ions 1Na + 2, for instance, which are found in higher concentrations outside a cell, the chemical force is directed inward. Thus an
outwardly directed electrical force is required to balance the chemical force. Because Na + is positively charged, a positive membrane
potential will exert an outward electrical force that balances the
inward chemical force. This means that the Na + equilibrium potential must be positive. If an ions concentration on either side of
a membrane is known, its equilibrium potential can be calculated
using the Nernst equation (Toolbox: Equilibrium Potentials and
the Nernst Equation).

+
Electrical driving force

Potencial electroqumico
1-Potencial qumico: a favor del
gradiente;
2-potencial elctrico: tendencia
de los iones de ser empujados
p a rDetermining
a u n the
l aDirection
d o uof theoElectrochemical
t r o
Driving Force The direction of the electrochemical driving
force acting on an ion
can be potencial
determined using thede
following prodependiendo
del
cedure. To carry it out, we need to know the ions equilibrium
potential (which is given the symbol E for ion X) and the cells
membrana
membrane potential:
X

1.

Vm = 100 mV
(b)

2.
Species

+
++

Vm = 50 mV
(c)

Using the principles described in the preceding sections,

identify the directions of the chemical and electrical driving
0
forces acting on the ion. If the two driving forces are going in
ion
the same direction, the electrochemical force also acts in that
i is necessary.
direction, and no further analysis

Electrical driving force

Na+

RT I
ln
zF I

If the chemical and electrical forces act in opposite directions,

compare the sizes of the equilibrium potential and the mem[Extracellular]
[Intracellular]
potential then the elecbrane potential.
If they are Equilibrium
equal in magnitude,
mM
mM
(V ) mV
trochemical force is zero and the ioneqis at equilibrium. If not,
proceed
to step 3.
150
15
+57.3

150
-81.2
If5.5
the equilibrium
potential is larger
in magnitude than the
membrane
potential,
1.5
0.0002then the chemical
+119.2 force is larger than the
electrical
force; the
force, therefore, acts in
Cl125
9 electrochemical
-70.3
the same direction as the chemical force. If the membrane potential is larger in magnitude than the equilibrium potential,
then the electrical force is larger than the chemical force; the
electrochemical
force, therefore,
in the
direction as
Because ECl = V (membrane
potential),
no forcesacts
other
thansame
those
the electrical force.
K+

Ca2+

3.

EnergYca potencial electroqumico

j = 0j + RT lnC j + Z j F

In
Nai+
G = GC + Gm = RT ln + + FE
Nao

Out (sangre)

Transporte Pasivo: cmo

m o d i c a r l a
permeabilidad de la
membrana para ciertas
molculas?

Difusin Facilitada por
Canales y/o Ionforos

Transporte Pasivo-Difusin facilitada

Energa de la molcula/
in en el interior de la
membrana
Energa de la molcula/in
en el interior de la
membrana en presencia del
ionforo/transportador/canal

Alta
concentracin

Baja
concentracin

Transporte Pasivo-Difusin facilitada

Regulacin de J:
Mediado por molculas que regulan la velocidad de difusin
espontnea (regulando la permeabilidad) y que presentan cierta
AFINIDAD y ESPECIFICIDAD por el sustrato

Molculas que modifican P de iones:

#

ula particin del ion a membrana por neutralizacin o

formando un ion de menor densidad de carga
u generando un entorno de mayor constante dielctrica

Transporte Pasivo-Difusin facilitada

Tipo de molculas que modifican la permeabilidad de
membranas:
u
u
u
u
u

Polipptidos cclicos (valinomicina, eniantina)

Polipptidos lineales (gramicidina)
Heterociclos con oxgeno (nonactina)
Politeres cclicos (alameticina)
Antibiticos cclicos (nistatina, anfotericina, filipina)

Molculas que ejercen su accin en MEMBRANAS

Transporte Pasivo-Difusin facilitada

H+, K+ anYporter

#
H+, Na+ anYporter

Grupos polares hacia el interior y

no polares al exterior

Complejos 1:1 NEUTROS

con metales M+

Anfotericina (poro,
Ergosterol)

Nonactina
(K+, NH3+)

Nistatina (poro;
Ergosterol)

Valinomicina (K+)

Transporte Pasivo-Difusin facilitada

Canales: protenas integrales

Transporte Pasivo-Difusin facilitada

Gramicidina A (Bacillus brevis)
Usado como antibitico

Pptido de 15 aa (LyD)
Alta especificidad para metales +1
Transporta 107 cationes por s
Cs+>Rb+>K+>Na+>Li+ (como el tamao y D)

Blanco: C
Azul: N
Rojo: O
H = 2.6 nm

Canal formado por un pptido en cada hemicapa

Transporte Pasivo-Difusin facilitada

Otros inoforos formadores de canales:
Pptidos que forman agregados de hlices anfipticas

Melitina (abeja)
Cecropina (gusano de seda)
Alameticina (hongo): controlado por
potencial

Transporte Pasivo-Difusin facilitada

Regulacin de canales:

Transporte Pasivo-Difusin facilitada

Receptor de acetilcolina:
Canal para cationes estimulado por neurotransmisor
Na+ es el ion que mas se transporta (tambin K+)
El canal se abre al unirse 2 acetilcolinas

Transporte Pasivo-Difusin facilitada

Canales sensibles al potencial:
Canales de sodio y de potasio activados por potencial
Un aumento abrupto en Na+, cambia el valor del potencial:
De -100 mVeqK a 70 mVeqNa : depolarizacin local de la
membrana.

Canal de sodio activado por

potencial
Selectivo a sodio (por tamao)
-100mV 5% del tiempo abierto
+70 mV 90%

Transporte Pasivo-Difusin facilitada

Transportadores

Uniporter: GLUT1-Glucosa

1. Vuniporter>>Vdif. Pasiva 2. Las molculas no se insertan en la membrana propiamentedicha (no hay particin). 3. Hay un nmero
definido de transportadores (Vmax, Saturable) 4. El transporte es especfico (Km).

Canales

Las caractersYcas ltrantes de cada canal se deben a:

1- las anidades de unin entre los sustratos (molculas/
iones que pasan a travs del canal)

2- el la geometra molecular (area de exclusin)

3- Las interacciones dinmicas entre el in y las paredes del
canal (difusin molecular en el canal)

4-La energa de hidratacin molecular o inica

Transporte AcYvo-Cotransporte o transporte acYvo secundario

Se transportan dos molculas a la vez donde una es a favor (ion) y la otra es en contra
(molcula orgnica pequea, azcares, iones) del gradiente de concentracin
La energa para el transporte contra-gradiente se obYene del gradiente electroqumico
de los iones Na+ o H+
Simporter

Equilibrio

An9porter

-6 Kcal

6000cal/mol

[glucosa]in
1
1
= e 1.987cal.mol .K 300 K = 23500
[glucosa]out

Para que haya transporte

pasivo y co-transporte activo
debe haber un gradiente:

Transporte AcYvo-Bombas acYvadas por ATP

Convierte energa qumica (hidrlosis de ATP) a mecnica para transportar
iones y pequeas molculas en contra del gradiente (electro)qumico.
Movimiento neto de una carga hacia fuera: Bomba Electrognica

Transporte AcYvo-Bombas acYvadas por ATP

Bomba Na+/K+
Mantiene gradiente (Na+ /K+) indispensable para el correcto funcionamiento
fisiolgico celular. Consume ms de 30% del consumo de ATP del animal en
reposo (en neurona 70% de la energa celular)
In

Species

[Extracellular]
mM

Na+

Out (sangre)

[Intracellular]
Species Equilibrium
[Extracellular]
potential
[Intracellular]
mM
mM
(Veq) mV
mM

150

15
Na+

150
+57.3

15

K+

5.5

150
K+

5.5
-81.2

150

Ca2+

1.5

0.0002
Ca2+

1.5
+119.2

0.0002

125

9Cl-

125
-70.3

Cl-

E l G p a r a e l
transporte de K al
interior celular es bajo
Because E , E and E ya
V,
Because
this
suggests
E el
, E that
and
other
E processes
V, this suggests
interv
que
gradiente
de
to exclude Na and Ca and to accumulate
exclude NaK.
and
These
Ca and
are to
transport
accumulate
processes and must be ACTIVE.
pprocesses
o tandemustnbe ACTIVE.
c i a l
+ FE + GATPADP
e l e c t r o s t Y c o e s
favorable

Because ECl = V (membraneBecause

potential),
EClno
=V
forces
(membrane
other than
potential),
those
no
+
represented by the chemicalrepresented
and electrical
bygradients
the chemical
(the and
electrochemic
electrical
gradient) need be invoked togradient)
explain the
need
distribution
be invoked
of to
Cl-explain
across the
the di
c
membrane.
membrane.
Na

Ginout (total) = RT ln

Na
Na

+
i
+
o

Ca

Na

Ca

Transporte AcYvo-Bombas acYvadas por ATP

Bomba Na+/K+
ATP4-+H2O+3Na+(in)+2K+(out) ADP3-+H2PO4- +3Na+(out)+2K+(in)
BOMBA ELECTROGNICA: pasaje de corriente a travs de la membrana

Hidrlisis de ATP y transporte ALTAMENTE acoplados.

El proceso se puede revertir: al revertir los gradientes se sintetiza ATP
Contra el gradiente de concentracin
y a favor del de potencial: WK 8 kJ
(2 moles)

2K+

2K+
ATP4-

3 Na+

Fuente de energa
E 55 kJ/mol

ADP33 Na+

Contra el gradiente de concentracin

y de potencial: WNa 35 kJ (3 moles)

Eciencia: W / E 0.8 (80%)

Transporte AcYvo-Bombas acYvadas por ATP

Bomba H+/K+
H+, K+ ATPasa Gstrica
pH estomacal: 0.8-1
pH mucosa gstrica 7.4
pH = 6.6
Hidrlisis de ATP: intercambia K+ por H+

1- K+ (estmago) K+ (mucosa)
H+ (mucosa) H+ (estmago)
2- K+ (mucosa) K+ (estmago)
Cl- (mucosa) Cl- (estmago)
HCl (mucosa) HCl (estmago)

NO ELECTROGNICA

Resmen Tipos de Transporte a travs de la membrana

Bibliogra>a
- Lodish y col (2003). Molecular Cell Biology
- Stein, W.D. (1986) Transport and Diusion across Cell Membranes. Academic Press
- Staneld C.L (2013) Principles of Human Physiology