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2.1 Introduction

Cell Biology, Roosevelt University, Fall 2009

Chapter 2

Transport of ions and small molecules

across membranes
By
Stephan E. Lehnart & Andrew R. Marks

2.4 Electrochemical gradients across the cell membrane generate the membrane potential

•  Cells maintain a negative resting membrane

potential with the inside of the cell slightly
more negative than the outside.

•  The membrane potential is a prerequisite for

electrical signals and for directed ion
movement across cellular membranes.
•  The membrane potential across a cell
membrane is due to:
–  an electrochemical gradient across a membrane
–  a membrane that is selectively permeable to ions

2.12 Action potentials are electrical signals that depend on several types of ion channels

•  Repolarization is shaped by transport of K+

Action Potentials
•  Action potentials enable rapid communication
between cells.
•  Na+, K+, and Ca2+ currents are key elements
of action potentials.
•  Membrane depolarization is mediated by the
flow of Na+ ions into cells through voltage-
dependent Na+ channels.
ions through several different types of K+
channels.
•  The electrical activity of organs can be
measured as the sum of action potential
vectors.
•  Alterations of the action potential can
predispose for arrhythmias or epilepsy.

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•  There are two principal types of membrane

transport proteins:
–  Channels
–  Carriers

2.2 Channels and carriers are the main types of membrane transport proteins

•  transporters 2.3 Hydration of ions influences their flux

•  pumps through transmembrane pores
•  Salts dissolved in water form hydrated ions.
•  The hydrophobicity of lipid bilayers is a barrier
to movement of hydrated ions across cell
membranes.
•  By catalyzing the partial dehydration of ions,
ion channels allow for the rapid and selective
transport of ions across membranes.
•  Dehydration of ions costs energy, whereas
hydration of ions frees energy.

2.5 K+ channels catalyze selective and rapid ion permeation

•  A K+ channel is a complex of four identical
Structural features of channel proteins subunits, each of which contributes to the
pore.
•  Multiple subunits in symmetrical
arrangements
•  Membrane-spanning helices
•  Central pore
•  Selectivity filter
•  Gating mechanisms

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2.5 K+ channels catalyze selective and rapid ion permeation 2.5 K+ channels catalyze selective and rapid ion permeation

•  The selectivity filter of K+ channels is an

evolutionarily conserved structure.

•  The K+ channel selectivity filter catalyzes

dehydration of ions, which:
–  confers specificity
–  speeds up ion permeation

2.6 Different K+ channels use a similar gate coupled to different activating or inactivating mechanisms.

•  The K+ channel gate is distinct from the

selectivity filter.

•  K+ channels are regulated by the membrane

•  Gating is an essential potential.
property of ion
channels.

•  Different gating
mechanisms define
functional classes of K+
channels.

2.7 Voltage-dependent Na+ channels are activated by membrane depolarization and translate electrical signals

•  The inwardly directed

Na+ gradient maintained by the •  Electrical signals at the cell membrane activate
Na+/K+-ATPase is required for the function of Na+ voltage-dependent Na+ channels.
channels.
•  The pore of voltage-dependent Na+ channels is
formed by one subunit, but its overall architecture is
similar to that of 6TM/1P K+ channels.

•  Voltage-dependent Na+ channels are inactivated by

specific hydrophobic residues that block the pore.

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•  Cell surface Ca2+ channels translate membrane

signals into intracellular Ca2+ signals. Muscle Contraction

•  The process of excitation-contraction

coupling, which is initiated by membrane
depolarization, controls muscle contraction.

•  Ryanodine receptors and inositol 1,4,5-

trisphosphate receptors are Ca2+ channels
that release Ca2+ into the cytosol from
intracellular stores

2.13 Cardiac and skeletal muscles are activated by excitation-contraction coupling 2.13 Cardiac and skeletal muscles are activated by excitation-contraction coupling

•  Intracellular Ca2+ release through ryanodine

receptors in the sarcoplasmic reticulum
membrane stimulates contraction of the
myofilaments.

•  Several different types of Ca2+ transport

proteins, including the Na+/Ca2+-exchanger
and Ca2+-ATPase are important for
–  decreasing the cytosolic Ca2+ concentration
–  controlling muscle relaxation

2.11 Selective water transport occurs through aquaporin channels

•  The aquaporin selectivity filter has three

Aquaporins
major features that confer a high degree of
•  Aquaporins allow rapid
selectivity for water:
and selective water –  size restriction
transport across cell –  electrostatic repulsion
membranes. –  water dipole orientation

•  Aquaporins are
tetramers of four
identical subunits, with
each subunit forming a
pore.

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Glucose transport
•  To cross the blood-brain
barrier, glucose is
transported across
endothelial cells of
small blood vessels into
astrocytes.
•  Glucose transporters
are uniporters
2.15 Symporters and antiporters mediate coupled transport
2.16 The transmembrane Na+ gradient is
essential for the function of many
transporters
•  The plasma membrane Na+
gradient is maintained by the
action of the Na+/K+-ATPase.
•  The energy released by
movement of Na+ down its
electrochemical gradient is
coupled to the transport of a
variety of substrates.
–  Ca2+
–  Mg2+
–  Cl-
–  glucose
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Symporters and antiporters mediate
coupled transport
•  Bacterial lactose permease functions as a symporter.
–  It couples lactose and proton transport across the
cytoplasmic membrane.
•  Lactose permease uses the electrochemical H+
gradient to drive lactose accumulation inside cells.
•  Lactose permease can also use lactose gradients to
create proton gradients across the cytoplasmic
membrane.
2.15 Symporters and antiporters mediate coupled transport
•  The mechanism of transport by lactose permease
likely involves inward and outward configurations.
–  They allow substrates to:
•  bind on one side of the membrane and to
•  be released on the other side
2.18 The Ca2+-ATPase pumps Ca2+ into
intracellular storage compartments
•  Ca2+-ATPases undergo a
reaction cycle involving
two major conformations,
similar to that of Na+/K+-
ATPases.
•  Phosphorylation of Ca2+-
ATPase subunits drives:
–  conformational changes
–  translocation of Ca2+ ions
across the membrane
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2.19 The Na+/K+-ATPase maintains the 2.20 The F1Fo-ATP synthase couples H+
plasma membrane Na+ and K+ gradients movement to ATP synthesis or hydrolysis
•  The Na+/K+-ATPase is a P-type ATPase that is •  The F1Fo-ATP synthase is a
similar to the Ca2+-ATPase and the H+-ATPase. key enzyme in oxidative
phosphorylation.

•  The Na+/K+-ATPase maintains the Na+ and K+ •  The F1Fo-ATP synthase is a

gradients across the plasma membrane. multisubunit molecular
motor.
–  It couples the energy
•  The plasma membrane Na+/K+-ATPase is released by movement of
electrogenic: protons down their
electrochemical gradient to
–  it transports three Na+ ions out of the cell for every two K+ ATP synthesis.
ions it transports into the cell.

2.21 H+-ATPases transport protons out of the cytosol

2.21 H+-ATPases transport protons out of •  V-ATPases in the
the cytosol plasma membrane
•  Proton concentrations affect serve specialized
many cellular functions. functions in:
–  acidification of
extracellular fluids
•  Intracellular compartments are
acidified by the action of V- –  regulation of cytosolic
pH
ATPases.

•  V-ATPases are proton pumps

that consist of multiple
subunits, with a structure
similar to F1Fo-ATP synthases.

3.1 Introduction

•  Cells must localize proteins to

specific organelles and
membranes.

•  Proteins are imported from the

cytosol directly into several types
of organelles.
Chapter 3
•  The endoplasmic reticulum (ER):
–  is the entry point for proteins
into the secretory pathway
Membrane targeting of proteins
–  is highly specialized for that
By purpose
D. Thomas Rutkowski & Vishwanath R. Lingappa
•  Several other organelles and the
plasma membrane receive their
proteins by way of the secretory
pathway.

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3.2 Proteins enter the secretory pathway by translocation across the ER membrane

•  Secretory proteins translocate completely across the ER

membrane;
–  transmembrane proteins are integrated into the membrane.

•  Signal sequences target •  Before leaving the ER, proteins are modified and folded by
nascent secretory and enzymes and chaperones in the lumen.
membrane proteins to the
ER for translocation.
•  The only feature common
to all signal sequences is
a central, hydrophobic
core that is usually
sufficient to translocate
any associated protein.

•  Proteins translocate through

•  Docking of SRP with its
receptor brings the
ribosome and nascent
chain into proximity with
the translocon.
•  Numerous accessory
•  Docking requires the
GTP binding and
hydrolysis activities of
SRP and its receptor.
an aqueous channel
composed of the Sec61
complex, located within the
ER membrane.
proteins that are involved in:
–  Translocation
–  Folding
–  Modification associate with the
channel

•  An interaction •  An interaction
between the between the
translocon and the translocon and the
signal sequence signal sequence
causes the channel causes the channel
to open and initiates to open and initiates
translocation. translocation.

•  The exact •  The exact

mechanism of mechanism of
translocation may translocation may
vary from one vary from one
protein to another. protein to another.

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3.8 Some proteins target and translocate posttranslationally

•  Posttranslational translocation proceeds 3.9 ATP hydrolysis drives translocation

independently of both ribosomes and SRP
•  The energy for
posttranslational
translocation comes from
ATP hydrolysis by the BiP
protein within the ER
lumen.

•  The energy source for

cotranslational
translocation is less clear,
but might be the same as
for posttranslational
translocation.

3.10 Transmembrane proteins move out of the translocation channel and into the lipid bilayer

•  Transmembrane domains exit the translocon

•  The synthesis of by moving laterally through a protein-lipid
transmembrane interface.
proteins requires
that trans-
membrane
domains be
–  recognized
–  integrated into the
lipid bilayer

•  Nascent chains are often

subjected to covalent
•  Oligosaccharyltransferase (OST) catalyzes N-linked
modification in the ER
lumen as they glycosylation on many proteins as they are translocated
translocate. into the ER.
•  The signal peptidase
complex cleaves signal
sequences.
•  GPI addition covalently
tethers the C-termini of
some proteins to the lipid
bilayer.

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•  Molecular chaperones associate with proteins in the

lumen and assist their folding. •  Protein disulfide isomerases (PDI) catalyze disulfide
•  Subunits that have not yet assembled into complexes are bond formation and rearrangement in the ER.
retained in the ER by interaction with chaperones.

3.19 Terminally misfolded proteins in the ER are returned to the cytosol for degradation

•  Calnexin and •  Proteins are returned to the cytosol by the

calreticulin escort process of retrograde translocation.
glycoproteins
through repeated
cycles of
chaperoning.
–  The cycles are
controlled by
addition and
removal of
glucose.

•  The major cellular

phospholipids are
synthesized predominantly
on the cytosolic face of the
ER membrane.
•  The localization of enzymes •  Each organelle has a unique composition of lipids.
involved in lipid –  This requires that lipid transport from the ER to
biosynthesis can be each organelle be a specific process.
controlled by the cell to
regulate the generation of
new lipids. •  The mechanisms of lipid transport between
•  Cholesterol biosynthesis is organelles are unclear.
regulated by proteolysis of –  They might involve direct contact between the ER
a transcription factor and other membranes in the cell.
integrated into the ER
membrane.
•  Transbilayer movement of lipids establishes
asymmetry of membrane leaflets.

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3.24 The ER is morphologically and functionally subdivided

•  The ER is morphologically subdivided into

specialized compartments, including:
–  the rough ER for protein secretion
–  the smooth ER for steroidogenesis and drug
detoxification
–  the sarcoplasmic reticulum for calcium storage
and release

•  The ER may also be subdivided at the

molecular level, in ways not morphologically
evident.

•  The extent and composition of the ER change in

response to cellular need.
Protein transport into other organelles

•  The ER moves along the cytoskeleton. •  Signal sequences are used for targeting to and
translocation across the membranes of other
organelles.

•  Mitochondria and chloroplasts are enclosed by a

double membrane, with each bilayer containing its
own type of translocon.

•  Two distinct pathways target matrix proteins to

peroxisomes.

•  Mitochondria have an inner and an outer membrane,

each of which has a translocation complex.
•  Import into mitochondria is posttranslational.
•  Mitochondrial signal sequences are recognized by a
receptor at the outer membrane.
•  The TOM complex (outer membrane) and
TIM complex (inner membrane) associate
physically, and the protein being imported
passes directly from one to the other.

•  Hsp70 (a chaperone) in the mitochondrial

matrix and the membrane potential across the
inner membrane provide the energy for
import.

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3.28 Complexes in the inner and outer membranes cooperate in mitochondrial protein import

•  Import into chloroplasts

occurs posttranslationally.

•  The inner and outer

membranes have
separate translocation
complexes that cooperate
during the import of
proteins.

•  Peroxisomal signal
sequences are:
–  recognized in the
cytosol
–  targeted to a
translocation channel

•  Peroxisomal proteins
are imported after they
are folded.

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