BISC 312 Lecture 7:

Lipids, Membrane, and Cellular Transport

Chapter 10
 Outline

 Molecular Structure of Lipids

• Fatty Acids
• Triacylglycerols

 Biological Membranes: Lipids

• Glycerophospholipids
• Sphingolipids and Glycosphingolipids
• Cholesterol

 Biological Membranes: Proteins and membrane structure

 Amphipathic properties of membrane lipids

• Apolar tail (hydrophobic) • Polar head (hydrophilic)

Polar head associates with water and nonpolar tail hides from water
 Lipids: Polar Fatty acids
Lipids are primary components of membranes

Lipids are amphipathic molecules:

• Polar head • Apolar tail (hydrophobic)

pKa= 4.5

Fatty acids are key components of lipids :long hydrocarbon chains (14-24 C) with carboxyl group

Stearate Saturated fatty acid

C18
18:0 (18C no double bound)

Oleic acid Unsaturated fatty acid

(lower melting point)
C18

18:1 (18C with 1 double bound)

Fatty acid tails: Unsaturated vs. Saturated

natural are almost

always in this form

Usually an even number of carbons

Unsaturation is important because it keeps membranes

fluid, instead of solid. Helps with movement and
conformation changes of transmembrane proteins

Straight fatty acid tails can pack together and interact

 Lipids: Fatty acids

Nomenclature: • Numbering starts from carboxyl group

• Last methyl carbon called ω (omega) carbon
• If double bond the end of the systematic name
changes from “…anoate” to “…enoate”
• Position of a double bond represented by a Δ (delta)
• A double bond can be cis or trans
 Neutral Fats (the fat we eat and store in body)

• Triacylglycerols:
fatty acids + glycerol

water-insoluble, so must have a specific mechanism of transport in body

 Effects of the amphipathic nature of membrane lipids
There is a preferential arrangement in micelles or bilayers to bury
hydrophobic fatty acids tails
• Single fatty acid chain: • Two fatty acid chains: formation of a
formation of a micelle bilayer

• Self assembly process driven by hydrophobic interactions between fatty acid chains

• Close packing is directed by van der Waals interactions between hydrocarbon chains
 What types of lipids occur in membranes?
• Most abundant: Glycerophospholipids

1 2 3
 Sphingolipids:
Differ from glycerol in C-2 hydroxyl is replaced by an –NH2 group and H on C-
1 is replaced by C15 hydrocarbon group
 Glycosphingolipids

• Glycolipids can have many carbohydrate residues

• Glycolipids are oriented asymmetrically in the plasma membrane
and are always facing the extracellular side.
Recall: can be components of RBC membranes
 Cholesterol in membranes
Acts as a fluidity buffer. Keeps membranes rigid at temps higher than
lipid melting point.

At lower temps it increases fluidity because it prevents close packing of

fatty acid structures

Weak amphipathic nature, polar group, rigid ring system, flexible tail
 Why so many lipids?
• They confer different properties on the membrane surface.

• Different cells have different membrane lipid composition: ie. brain cells
commonly have cerebrosides and gangliosides, axons are rich in
glycosphingolipids.

• Glycolipids are always on outer plasma membrane towards aqueous

environment. This is maintained by limited “flip-flop” of proteins.

• Some function as intracellular signaling molecules or interact with

proteins to to regulate activities of the cell.

• Some are of medical importance: gangliosides are not broken down

properly in Tay-Sachs disease or Gaucher’s disease, causing severe brain
disorders.
 Proteins are key components of membranes
Lipids provide barrier and proteins provide most functions of membranes
• Membrane proteins involved in:
-Transports across membranes
-Signaling and transfer of information (receptors)
-Maintenance of electric potential
• Depending on the membrane, lipid : protein ratio varies from 1:4 to 4:1

• Large variations in protein repertoire between

different membranes.

• Type of membrane proteins is related to

membrane function
Gel electrophoresis of protein content in:
A- Erythrocyte membrane (ABO group)
B- Retinal rod cells (photoreception)
C- Sarcoplasmic reticulum of muscle cells (Ca2+
storage and release)
Membranes and membrane proteins
Inserted into membranes as they
are synthesized by the cell

Lipid Fluid Mosaic Model:

How proteins and lipids interact in a
bilayer.
Free movement laterally

I = integral protein, present in bilayer

P = Peripheral protein, can be

associated by H-bonds
or ionic interactions.

A = Anchored protein, covalently

linked to fatty acid chains or
glycolipids at surface
 Diffusion depends on lipid composition

• Diffusion is mainly lateral and rapid, rarely transverse (flip-flop only for lipids)

• Membrane fluidity is controlled by fatty acid saturated unsaturated

composition and their melting temperature (Tm) to
semi-fluid state.

• Unsaturated fatty acids = less tight packing = disordered state

• Saturated fatty acids = tight packing = ordered state

• Length of fatty acid and degree of saturation affect

the overall melting temperature of membranes
 Diffusion depends on lipid composition (continued)

• Saturated lipids = tight packing = ordered/solid state

• Unsaturated = less tight packing = disordered/fluid state
Tm: transition temperature from a solid to fluid state

• Changing temperature can affects

membrane fluidity
• Reversibly, membrane can adapt to
changing temperatures by incorporating lipids
with different properties (more or less
saturated)
Bacteria and some hibernating animals adjust
degree of unsaturated fatty acids in their
membranes to deal with temperature
changes.
• Cholesterol is also an important regulator of
membrane fluidity by disrupting tight packing
 Membranes are asymmetric
Outer and inner membrane differ in lipid
composition and in types of associated proteins

• Asymmetry is mainly fixed at membrane

biosynthesis

• Asymmetry is maintained because rare flip-flopping

• Flippase can move lipids between inner and outer By Adam Steinberg

membranes (and vice versa)

• Orientation of transmembrane proteins

does not change overtime (important to
maintain membrane potential)

• Outer plasma membrane rich in

glycosylated protein and lipids
Integral Membrane Proteins

Held by noncovalent forces

Model of Glycophorin, major component

of red blood cell membrane
This protein is amphipathic

Extracellular portion would have hydrophilic

amino acids: ser, thr, asp, etc.

α-helix has many hydrophobic amino acids

isoleucine, leucine, valine, etc.

This protein is also a glycoprotein, meaning

it has carbohydrates (sugars) attached to it.
 Mechanism of protein/membrane interactions
Integral membrane proteins use different strategies to accommodate the polar
peptide backbone across the hydrophobic membrane core

- very common
• Alpha helices
- 7-span proteins
(e.g. G-coupled receptors) α-helices are formed
- 20 aa/helix of hydrophobic aa
- Extensive H-bonding residues

1 helix 3.6 aa/turn

1 turn = 0.54 nm
Bilayer core = 3 nm
aa/helix : ~20

Bacteriorhodopsin
 Functions of membranes
(plasma membrane)

• transport of substance in and out of cell

– Active vs. Passive
• ion transport and nerve cell conductance
• cell signaling (more later in the semester)
• maintaining shape
• cell-cell interaction
 Transport across membranes
Ability of a molecule to cross a membrane depends on:
1- The membrane permeability to the molecule
2- The presence of appropriate transporter and an energy source

For uncharged membrane permeable molecules chemical concentration gradient

determines spontaneous movement across membrane

ΔGTrans = RT ln (c2/c1) R: gas constant

[mol]
[mol]
T: temperature

For charged molecules one needs to consider an additional electrical potential term

ΔGTrans = RT ln (c2/c1) + ZF ΔV
Z: charge of the molecule ΔV: charge gradient
[Na+]
[Na+]
F: Faraday constant (potential for all ions) -+
----- + +
- + For all ions
+
ΔGTrans indicates if the transport is passive (ΔGTrans< 0) or if it requires energy
input (e.g.ATP) and is thus active (ΔGTrans> 0)
 Lipid bilayers are barriers to diffusion

• Low permeability to ions (need desolvation to go through the membrane directly)

• Low permeability to polar molecules (except H2O, small and uncharged)
• High selective permeability barriers to ions, using protein pumps and channels
- Maintain different ion conc. on each side of the membrane
(important for membrane potential)
 3 classes of proteins for transports across membranes

1- Membrane carriers/ • For passive transport or facilitated diffusion

Membrane channels • Provide a selective pore through which ion can flow rapidly
• When ΔGTrans< 0
• No use of ATP
• Can be sensitive to membrane polarization

2- Membrane pumps • For primary active transport

• When ΔGTrans> 0
• Drive thermodynamic uphill reactions with ATP as
free energy source
• Different pumps use different strategy for transport
and use of ATP

3- Membrane cotransporters • For secondary active transport

• When ΔGTrans> 0
• Drive thermodynamic uphill reactions using the
chemical gradient of one molecule to drive the transport
of a second molecule against its own gradient
• No use of ATP
 Membrane Channels
Permits the movement of ions
through the membrane down
concentration gradient
called facilitated diffusion or passive
transport.

ΔGTrans< 0, No energy required, but

allows charged molecules to move
across lipid bilayer.
 Aquaporin: A Membrane Channel

Aquaporin • Channels specialized in H2O transport

: • 6 α-helices forming hydrophilic
channel
• 106 H2O molecule/sec
• Important for rapid H2O transport
(e.g. reabsorption of water in kidney,
tears).
 Membrane ion channels

• Fastest transporters: 1000 fold faster than pumps

• Close to diffusion rates of ions
• Different ion channels use similar transport mechanisms yet are highly
selective for specific ions
• Channels can work because of ion concentration gradients generated by
membrane pumps in live cells

• Na+ and K+ ion channels are particularly important for signal

communication in the nervous systems
• Sequential triggering of Na+ and
K+ ion channels generates action
potential (nerve impulse)

Na+ ion channel K+ ion channel

Payandeh et al. Nature 2011, 475,353–358
 Structure of K+ channel reveals ion specificity
K+ channel is a tetramer (4 identical subunits) forming a pore through lipid
membranes

Prokaryotic K+ channel
 Structure of K channel reveals ion specificity (continued)
• Desolvated K+ ion interact with a specific aa sequence along the channel that provides
high selectivity for K+

• Peptidic backbone of 5 aa: TVGYG

from 2 subunits provides polar
interactions with K+ as a replacement
for H2O.
• Polar interaction compensate the
energy cost of desolvation for K+ but
not for other ions (e.g. Na+)
• Size of the pore and position of
TVGYG aa optimal for K+ ion only
(tight binding)

• Electrostatic repulsions between K+ ion in

channel allows for rapid flow
Gated ion-channel (passive transport system)

Gated pores that

respond to highly-
specific signals

Most important are

those for Na+, Ca2+,
and K+.

Passage of ions goes

down concentration
gradient.
Thus they are much
faster (1000x) than
transport
by Na+/K+ ATPase pump.

ie. Nerve impulse signals

 Voltage gating in K+ channels
Voltage gating is provided by a positively charged domain at the bottom of the
channel structure.

• S4++ domain form voltage

sensing paddles that can
occlude the channel

• Close channel: S4++ domain

“down position”

• On membrane
depolarization inside become
+++, electrostatic repulsion of
S4++ paddles upward: channel
opens
 Membranes pumps (Active Transport)
Involved in primary active transport of molecules (often ions) against their conc.
gradient. Need an energy source.

• ATP often the source of energy for this process

• Pumps have ATPase activity (hydrolyze ATP) to
transfer phosphate group to their own residues
(e.g. Asp)
• Free energy of ATP induces large
conformational changes in pumps (both
phosphorylation/dephosphorylation)

• Interconversion between 2
conformations (open/close) provides
unidirectional pumping of ions

• Pumps are important to maintain steady state concentration of ions in cells

- Regulation of Ca2+ conc. (important for cell signaling)
- Regulation of Na+/K+ conc. (important for membrane potential)
 Active Membrane Transport (Na+/K+ ATPase):
“P-Type ATPase”
Requires work, usually
provided by hydrolysis
of ATP.

Necessary to move substances

across concentration gradient

Animal cells have high

internal K+ and low
internal Na+ conc.
1) Protein (4 subunits) is open
to interior of cell and binds Na+
ATP phosphorylates protein.

2) conformational change. Open

to outside, doesn’t bind Na+, but
does bind K+. P is hydrolyzed, giving Pi

3) Goes back to original form, no longer

binds K+, which enters the cell. More
Na+ can now bind.
 Membrane carriers or Co-transporters
Involved in secondary active transport of ions. Use concentration gradient rather
than ATP for conformational changes
Secondary active transport: coupling a thermodynamically unfavorable reaction (e.g.
transport against conc. gradient) with a favorable one ( e.g. transport with conc. gradient)

COTRANSPORT
Antiporter Symporter Uniporter
Transport of 2 solutes in Transport of 2 solutes in Transport of a single solutes
opposite directions same direction following conc. gradient
 Cotransport systems
• The flow of ions down a gradient can be used to perform work.

• The high concentration of Na+ from outside the cell can flow back inside and
bring glucose along with it when BOTH are present in lumen of intestine. It
can transport glucose against concentration gradient by using potential
energy of Na+ gradient.