ChE 471

BIOCHEMISTRY

Lipids and Biological Membranes

Md. Burhan Kabir Suhan
Lecturer, Department of Chemical Engineering, BUET

Email: suhan@che.buet.ac.bd

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Special Courtesy: Mesbah Ahmad
Lipids
• lipids are a chemically diverse group of compounds with insolubility
in water being their defining feature.

• Fats and oils store energy

• Phospholipids and sterols are major structural elements of biological
membranes.
• Also acts as enzyme cofactors, electron carriers, light-absorbing
pigments etc.

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Storage Lipids
Fatty Acids
• The fats and oils used almost universally as stored forms of energy in living
organisms are derivatives of fatty acids.
• Long chain carboxylic acids
• Hydrocarbon chains ranging from 4 to 36
• Some have unbranched chains and are fully saturated while others have
double bonds and may contain three-carbon rings, OH group etc.
• By nature, fatty acids are AMPHIPATHIC – have both hydrophilic and
hydrophobic parts.
• The cellular oxidation of fatty acids (to CO2 and H2O), like the controlled,
rapid burning of fossil fuels in internal combustion engines, is highly
exergonic.

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Fatty Acids (contd)

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Fatty Acid Nomenclature

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Fatty Acid Nomenclature

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Fatty Acid Nomenclature
Draw 16:1(Δ9) i.e. trans-9-Hexadecenoic acid also known as Palmitoleic
acid.

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Fatty Acid Nomenclature

Draw 16:1(Δ9) i.e. trans-9-Hexadecenoic acid also known as

Palmitoleic acid.

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Fatty Acids (contd)
• In most monounsaturated fatty acids the double bond is between C-9
and C-10 (Δ9)
• The other double bonds of polyunsaturated fatty acids are generally
Δ12 and Δ15
• The double bonds of polyunsaturated fatty acids are almost never
conjugated (alternating single and double bonds) but are separated by
a methylene group.

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Fatty Acids (contd)
In nearly all naturally occurring unsaturated fatty acids, the double bonds are in the
cis configuration.
Trans fatty acids are produced by fermentation in the rumen of dairy animals and
are obtained from dairy products and meat. Because diets high in trans fatty acids
correlate with increased blood levels of LDL (bad cholesterol) and decreased HDL
(good cholesterol), it is generally recommended that one avoid large amounts of
these fatty acids. Unfortunately, French fries, doughnuts, and cookies tend to be
high in trans fatty acids.

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Fatty Acids (contd)
• The non-polar hydrocarbon chains account for the poor solubility of
FAs in water.

• The longer the fatty acyl chain and the fewer the double bonds, the
lower is the solubility in water.

• The carboxylic acid group is polar (and ionized at neutral pH) and
accounts for the slight solubility of short-chain fatty acids in water.

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Fatty Acids (contd)
• At room temperature, the saturated fatty acids from 12:0 to 24:0
have a waxy consistency, whereas unsaturated fatty acids of these
lengths are oily liquids.

• This difference in melting points is due to different degrees of packing

of the fatty acid molecules

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Fatty Acids (contd)
•
Fully saturated fatty acids
in the extended
form pack into nearly
crystalline arrays,
stabilized by many
hydrophobic interactions

The presence of one

or more cis double
bonds interferes
with this tight
packing and results
in less stable
aggregates

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Triacylglycerols (TAGs)
• Simplest lipids constructed from fatty acids
• Also referred to as triglycerides, fats and natural fats.
• Triacylglycerols are composed of three fatty acids each in ester
linkage with a single glycerol

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• the polar hydroxyls of glycerol and the polar carboxylates of the fatty
acids are bound in ester linkages, triacylglycerols are nonpolar,
hydrophobic molecules, essentially insoluble in water.

• Lipids have lower specific gravities than water, which explains why
mixtures of oil and water have two phases: oil, with the lower specific
gravity, floats on the aqueous phase.

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• In most eukaryotic cells, triacylglycerols
form a separate phase of microscopic,
oily droplets in the aqueous cytosol,
serving as depots of metabolic fuel.
• In vertebrates, specialized cells called
adipocytes, or fat cells, store large
amounts of triacylglycerols as fat
droplets (a) that nearly fill the cell.
• In some animals, triacylglycerols stored
under the skin serve not only as energy
stores but as insulation against low
temperature .

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Triacylglycerols (contd)
• Enzymes called lipases can break fats back to their constituent
species

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Biological waxes
• Biological waxes are esters of long-chain (C14 to C36) saturated and
unsaturated fatty acids with long-chain (C16 to C30) alcohols.
• Their MPs (60 to 100°C) are generally higher than those of
triacylglycerols.
• Water-repellent and have firm consistency.
• Have variety of applications in pharmaceuticals, cosmetics etc.
Lanolin (from lamb’s wool), beeswax, carnauba wax (from a Brazilian palm
tree), and wax extracted from spermaceti oil (from
whales; are widely used in the
manufacture of lotions, ointments,
And polishes.

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Structural Lipids in Membranes
• Biological membrane is a double layer of lipids.
• Membrane lipids are amphipathic: one end of the molecule is
hydrophobic, the other hydrophilic (polar head group and non-polar
tail).
• The properties of head groups determine the surface properties of
membranes.
• Different organisms and tissues have different membrane lipid head
group compositions.

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Glycerophospholipid

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Glycerophospholipid (contd)
• Similar in structure to triacylglycerols except one of the alcohols of
glycerol is esterified by phosphoric acids instead of a fatty acid

• Unsaturated fatty acids are commonly found connected to C2

• In general, glycerophospholipids contain a C16 or C18 saturated fatty
acid at C-1 and a C18 to C20 unsaturated fatty acid at C-2

Example- Phosphatidic acid (X=H)

Phosphatidylethanolamine (X= Ethanolamine)

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Glycerophospholipid (contd)
• Glycerol itself is not chiral, as it has a plane
of symmetry through C-2.

• However, glycerol can be converted to a

chiral compound by adding a substituent
such as phosphate to either of the CH2OH
groups; that is, glycerol is prochiral

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Glycerophospholipid (contd)
• Phospholipids are MUCH MORE
amphiphilic than triacylglycerols
due to CHARGED groups at neutral
pH
• Has both hydrophilic and
hydrophobic regions
• Therefore we can say that
phospholipids have:
– One POLAR HEAD
– TWO NON-POLAR TAILS

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Sphingolipids

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Sphingolipid (contd)
• Membrane lipids based on the core structure of SPHINGOSINE, a
long chain amino alcohol

• Glycerol is replaced by sphingosine

• A fatty acid is joined to sphingosine via an amide linkage rather than
an ester linkage as usually seen in lipids
• A polar head group is connected to sphingosine by a glycosidic or
phosphodiester linkage
• The sugar-containing glycosphingolipids are found largely in the outer
face of plasma membranes

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Sphingolipid (example)
- Ceramide (sphingosine + amide-linked fatty acid)
- Ceramide is the structural parent of all sphingolipids.
- three subclasses of sphingolipids, differing in their head groups:
sphingomyelins, neutral (uncharged) glycolipids, and gangliosides.
• Sphingomyelin
- Sphingomyelin is abundant in myelin sheath that surrounds
some nerve cells in animals

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Galactolipid and Sulfolipid

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Galactolipid (contd)
• One or two galactose residues are connected by a glycosidic
linkage to C-3 of a 1,2- diacylglycerol.
• Galactolipids are localized in the thylakoid membranes (internal
membranes) of chloroplasts;
• They make up 70% to 80% of the total membrane lipids of a vascular
plant.
• They are probably the most abundant membrane lipids in the
biosphere.
• Plant membranes also contain sulfolipids, in which a sulfonated
glucose residue is joined to a diacylglycerol in glycosidic linkage.

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Archaebacterial ether lipids

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Archaebacterial ether lipids (contd)
• The archaebacteria have membrane lipids containing long chain (32
carbons) branched hydrocarbons linked at each end to glycerol.

• These linkages are through ether bonds, which are much more stable
to hydrolysis at low pH and high temperature than are the ester bonds
found in the lipids of eubacteria and eukaryotes.

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 Sterols
• Sterols are structural lipids present in the membranes
of most eukaryotic cells
• Based on a fused ring system
– RIGID structure
• Includes HORMONES (testosterone,
progesterone, estrogen), cholesterol

Steroid nucleus:
• Four fused rings
• Hydroxyl group (polar head) in
the A-ring
• Various nonpolar side chains
• The nucleus is almost planar
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Sterols (contd)
• Cholesterol and related sterols are present in the membranes of most
eukaryotic cells
– Thicken the plasma membrane
– maintain membrane fluidity
– Most bacteria lack sterols
• Mammals obtain cholesterol from food or synthesize it in the liver
• Bacteria cannot synthesize sterols; a few bacterial species, however,
can incorporate exogenous sterols into their membranes

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Steroids
-Contain the gonane structure (tetracyclic hydrocarbon with no double
bonds), additional functional groups, and/or modified ring systems
derived from gonane
- Sterols are steroid alcohols with an OH group
- More polar than cholesterol
- Steroid hormones are synthesized from cholesterol in gonads and
adrenal glands
- They are carried through the body in the bloodstream, usually
attached to carrier proteins
- Many of the steroid hormones are male and female sex hormones

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Steroids

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Lipids as signals, cofactors and Pigments
• Compounds with long conjugated systems absorb light in the
visible region of the spectrum.
Subtle differences in the chemistry of these compounds
produce pigments of strikingly different colors.

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 Biological membranes
• A lipid bilayer is the basic structural element of membranes that surround all
cells and organelles

- Made up of phospholipids, glycosphingolipids, sphingolipids

and cholesterol (if animal)
- Non-polar components minimize
exposure to water
by forming a bilayer
• Polar head groups face
outward and H-bond with
water
• Lipid fatty acid chains face
inward and interact via
hydrophobic interactions

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Membrane composition
• Each Type of Membrane Has
Characteristic Lipids and Proteins
• The relative proportions of
protein and lipid vary with the
type of membrane
• In all but a few cases, the
functional significance of these
combinations is not yet known.
• Cholesterol is prominent in plasma membranes
but barely detectable in mitochondrial
membranes.
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 Membrane composition (contd)
However, all biological membranes share
some fundamental properties:
1. Membranes are impermeable to most
polar or charged solutes, but permeable to
nonpolar compounds
2. They are 5 to 8 nm (50 to 80 Å) thick
3. Appear trilaminar when viewed in cross
section with the electron microscope
4. The trilaminar image consists of two electron-
dense layers (bound to the inner and outer
surfaces of the membrane) separated by a less
dense central region

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How are membrane bilayers formed?
• Phospholipids spontaneously form
stable bilayers, with their polar head
groups exposed to water and their
hydrophobic tails buried in the interior
of the membrane.
• It is the shape and amphipathic nature of
the lipid molecules that cause them to form
bilayers spontaneously in aqueous
environments.
• Lipids constitute approximately 50% of the
mass of most cell membranes, although
this proportion varies depending on the
type of membrane. 46
Lipid aggregates
Depending on the precise conditions and the nature of the lipids,
three types of lipid aggregates can form when amphipathic lipids
are mixed with water

Liposome
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Micelle
• Micelles are spherical structures
• Forms in the solution of amphipathic
molecules that have larger head than tail
– Fatty acids
– Sodium dodecyl sulfate
• Each micelle has from a few
dozen to a few thousand lipid
molecules
• Aggregation occurs when the
concentration of molecules is
higher than a certain threshold

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Bilayer
• Consists of two lipid monolayers
• Occurs most readily when the cross-
sectional areas of the head group and
acyl side chain(s) are similar
• Hydrophilic head groups interact
with water
• Hydrophobic fatty acid tails are
packed inside
• One leaflet faces the cytoplasm
• Another leaflet faces the
extracellular space

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Liposome
• Bilayer sheet is relatively unstable and spontaneously
forms Liposome by folding back on itself to form a
hollow sphere
• By forming vesicles, bilayers lose their hydrophobic
edge regions, achieving maximal stability in aqueous
environment
• These bilayer vesicles enclose water, creating a
separate aqueous compartment.
• The central aqueous cavity can enclose dissolved
molecules
• They are useful artificial carriers of molecules (e.g.,
drugs)
• Vesicles fuse readily with cell membranes or with each
other

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Membrane properties
1. Stable: once formed it stays formed and unlikely to disintegrate
– Due to weak interactions among nonpolar tail regions
– tendency of highly dipolar water molecules to exclude nonpolar
molecules from their midst.
2. Prevents passage of polar substances
– forms a boundary and compartmentalizes regions of cytoplasm
containing relatively water-soluble and nonamphipathic solutes , such
as ions , sugars, amino acids , and nucleotides and much larger
molecules, such as proteins , that are unable to pass easily across it

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Membrane properties (contd)
3. Dynamic nature
– A membrane will bend and fold; it can be deformed without breaking
– If broken, it very quickly reseals, e.g vesicle fusion or budding
– At room temperature a membrane's constituents are in constant,
random motion.
– The amphipathic nature of its lipid constituents makes a membrane
both a stable and a fluid boundary.

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Fluid mosaic model
• Cell membrane behaves as a two-dimensional liquid in which
phospholipid and protein molecules diffuse easily
• Proteins and substances such as cholesterol become embedded in the
bilayer, giving the membrane the look of a mosaic.
• Integral proteins are firmly associated with the membrane, often
spanning the bilayer
• Peripheral proteins are weakly associated and can be removed easily
– Some are noncovalently attached
– Some are linked to membrane lipids

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• the plasma membrane has the consistency such that the proteins and
other substances are able to move across it. That’s why it’s called the
fluid-mosaic model.

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 Peripheral Membrane Proteins
• Relatively loosely associated with
membrane
-- Associated with the polar head groups
of membranes
– through ionic interactions and hydrogen
bonding with the hydrophilic domains of
integral membrane proteins
• can be released by disrupting ionic
interactions or break H bonds either with
change in ionic strength or change in pH
• Peripheral proteins may serve as
regulators of membrane-bound enzymes
or may limit the mobility of integral
proteins by tethering them to intracellular
structures
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Integral Membrane Proteins
• Span the entire membrane
• Have asymmetry like the membrane
– Different domains in different
compartments
• Tightly associated with membrane
– Hydrophobic segment in the protein
interacts with the hydrophobic regions of the
membrane
• Extraction of membrane proteins from
biological membranes is usually accomplished
with the help of detergents
• Purified integral membrane proteins still
have phospholipids associated with them
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Integral Membrane Proteins (contd)
Types I and II have only one transmembrane helix; the amino-terminal domain is outside
the cell in type I proteins and inside in type II.
Type III proteins have multiple transmembrane helices in a single polypeptide.
In type V proteins, transmembrane domains of several different polypeptides assemble to
form a channel through the membrane.
Type IV proteins are
held to the bilayer
primarily by
covalently linked
lipids
Type VI proteins
have both
transmembrane
helices and lipid
glycosylphosphatidylinositol
(GPI) anchors.

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 GPI anchored proteins

Structure of the GPI anchor from human erythrocyte

acetylcholinesterase. The three domains of the GPI
anchor are (i) a phosphoethanolamine linker (red), (ii)
the conserved glycan core (black), and (iii) a
phospholipid tail (blue). Appendages in blue (including
the lipids of the lipid tail) are variable.

Without GPI anchored proteins

linked to the cell surface,
the complement
system can lyse the cell, and high
numbers of RBCs are destroyed,
leading to hemoglobinuria
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GPI anchored proteins

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Membrane dynamics
Motion of single phospholipids in a
bilayer:
(a) Uncatalyzed movement from
one leaflet to the other is very
slow (requires a large free
energy change)
(b) A family of proteins, the flippases,
facilitates flipflop diffusion
(c) lateral diffusion within the
leaflet is very rapid, requiring
no catalysis
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Membrane dynamics (contd)
• Individual lipid molecules can move laterally in the plane of the
membrane by changing places with neighbouring lipid molecules.

• A molecule in one monolayer, or leaflet, of the lipid bilayer - the outer

leaflet of the erythrocyte plasma membrane, for example can diffuse
laterally so fast that it circumnavigates the erythrocyte in seconds.

• This rapid lateral diffusion within the plane of the bilayer tends to
randomize the positions of individual molecules in a few seconds

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Membrane dynamics (contd)
Lateral diffusion can be shown experimentally by
attaching fluorescent probes to the head groups of
Lipids and using fluorescence microscopy to follow
the probes over time.

Measurement of lateral
diffusion rates of lipids by
fluorescence recovery after
photobleaching (FRAP)
• The rate of fluorescence recovery
after photobleaching, or FRAP, is
a measure of the rate of lateral
diffusion of the lipids.
• Using FRAP technique,
researchers have shown that
some membrane lipids diffuse
laterally by up to 1 µm/s
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Membrane transport
Transport through a biological membrane can
take place via the following processes:
1. Simple diffusion
2. Facilitated diffusion
3. Primary active transport
4. Secondary active transport
5. Ion channel
6. Ionophore-mediated ion transport

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Simple diffusion
• During simple diffusion, a molecule
simply dissolves in the phospholipid
bilayer, diffuses across it, and then
dissolves in the aqueous solution at the
other side of the membrane.
• No membrane proteins are involved
• The direction of transport is determined
simply by the relative concentrations of
the molecule inside and outside of the
cell.
• Importantly, only small relatively
hydrophobic molecules are able to diffuse
across a phospholipid bilayer at significant
rates.

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Simple diffusion

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When ions of opposite charge are separated by a permeable membrane, there is a
transmembrane electrical gradient, a membrane potential, Vm (expressed in volts or
millivolts).
Thus the direction in which a charged solute tends to move spontaneously across a
membrane depends on both the chemical gradient (the difference in solute
concentration) and the electrical gradient (Vm) across the membrane. Together,
these two factors are referred to as the electrochemical gradient or electrochemical
potential. 69
Facilitated diffusion
Passage is mediated by proteins that enable
the transported molecules to cross the
membrane without directly interacting with
its hydrophobic interior.
Two classes of proteins that mediate facilitated
diffusion are generally distinguished: carrier
proteins and channel proteins.
• Carrier proteins bind specific molecules
to be transported on one side of the
membrane. They then undergo
conformational changes that allow the
molecule to pass through the membrane
and be released on the other side.
• In contrast, channel proteins form open
pores through the membrane, allowing
the free diffusion of any molecule of the
appropriate size and charge. 70
Facilitated diffusion

Ref: "Scheme facilitated diffusion in cell membrane" by Mariana Ruiz Villareal

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Regulation by insulin of
glucose transport by GLUT4
into a myocyte.

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Primary and Secondary active transport

a) In primary active transport, the energy released by ATP hydrolysis drives solute movement against an
electrochemical gradient. (b) In secondary active transport, a gradient of ion X (often Na) has been
established by primary active transport. Movement of X down its electrochemical gradient now provides the
energy to drive cotransport of a second solute (S) against its electrochemical gradient.
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Energy requirement
The amount of energy needed for the transport of a solute against a gradient
can be calculated from the initial concentration gradient. The general equation
for the free-energy change in the chemical process that converts S to P is:
∆G= ∆G’o + RT ln [P]/[S]
Where, R is the gas constant, 8.315 J/mol.K, and T is the absolute temperature.

When the “reaction” is simply transport of a solute from a region where its
concentration is C1 to a region where its concentration is C2, no bonds are
made or broken and the standard free-energy change, ΔG’0, is zero. The free-
energy change for transport, ΔGt, is then
𝐶2
ΔGt= RT ln
𝐶1

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Energy requirement

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Energy requirement (contd)
When the solute is an ion, its movement without an accompanying
counterion results in the endergonic separation of positive and negative
charges, producing an electrical potential; such a transport process is said to
be electrogenic. The energetic cost of moving an ion depends on the
electrochemical potential, the sum of the chemical and electrical gradients:

where Z is the charge on the ion, F is the Faraday constant (96,480 J/V.mol),
and Δψ is the transmembrane electrical potential (in volts).
Eukaryotic cells typically have electrical potentials across their plasma
membranes of about 0.05 to 0.1 V

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Energy requirement (contd)

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Energy requirement (contd)

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 Ionophores
Ionophores are lipid-soluble molecules that bind specific ions and carry
them passively across membranes, dissipating the energy of
electrochemical ion gradients.

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• Because of the essential role of ion gradients in active transport and
energy conservation, compounds that collapse ion gradients across
cellular membranes are effective poisons, and those that are specific
for infectious microorganisms can serve as antibiotics. One such
substance is valinomycin, a small cyclic peptide that neutralizes the K+
charge by surrounding it with six carbonyl oxygens . The hydrophobic
peptide then acts as a shuttle, carrying K+ across membranes down its
concentration gradient and deflating that gradient. Compounds that
shuttle ions across membranes in this way are called ionophores (“ion
bearers”). Both valinomycin and monensin (a Na+ carrying ionophore)
are antibiotics; they kill microbial cells by disrupting secondary
transport processes and energy conserving reactions.
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Ionchannel

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Summary
of Transport
Types

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END

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