Chapter 10
Lipids
White Fat:
Adipocytes
Lipids Fall Into 3 Functional Groups
Storage Lipids
Contain Fatty acids and oils
Built to make them stable (i.e. inert)
and easy to disassemble
Structural Lipids
Contain Phospholipids and sterols
Built to provide stability and mobility
for membrane components
Others
Contain cofactors, electron carriers,
pigments, hydrophobic anchors,
hormones, intracellular messengers
Built for specialized functions
Lipid Structural Features
Backbones
- Sphingosine
Glycerol
Fatty Acids
- Hydrocarbons
(R-Groups)
Complex rings
- Cholesterol and derivatives
- Cofactors
- Signaling molecules
- Vitamins
STORAGE LIPIDS: Fatty Acids and Glycerol
Fatty acid Structure
Hydrocarbon Derivatives
Carboxylic acids with hydrocarbon chains
May be
~ 4-36 carbons
- Fully or partly saturated
- Branched or unbranched
May contain
- Three-carbon rings,
- R-groups:
and
methyl, hydroxyl
Biological fatty acids do not usually
Double bonds are usually are
have conjugated double bonds
separated by a methylene (group)
-CH=CH-CH=CH-
-CH=CH-CH2-CH=CH-
Fatty Acid Nomenclature
Nomenclature
Based on chain length : double bond character
Palmitic acid
Oleic acid
16:0
18:1
Position of double bond is identified:
20 carbon chain with 2 double bonds:
15
20:2( 9,12)
9
Monounsaturated (one double bond)
Polyunsaturated (more than one double bond)
12 and
The most common fatty acids: 12-24 carbons, unbranched chains
- Even number of carbons
- Acetate condensation (two carbons) = even number
STORAGE LIPIDS
Physical Properties
Chain is non-polar
- Fatty acids tend to be poorly water-soluble
- Slight solubility in water is due to carboxyl groups
At room temperature
- Saturated fatty acids are waxy solids
- Unsaturated fatty acids are more liquid, oily
What chemical properties cause these differences?
Saturated chains
are linear
Unsaturated chains
contain bends
Triacylglycerols: Fatty Acid Esters of Glycerol
Glycerol + 3 fatty acids
Ester bonds
Hydrophobic, nonpolar
Nearly insoluble in water
The storage form
of both glycerol
and fatty acids
Cleaved by Lipases:
hydrolyze ester linkages
Advantages of using triacylglycerols as fuels
1. Hydrophobic No solvation layer for each molecule
- One layer around many molecules
Saves energy
2. Carbon atoms are highly reduced
- Yield more energy than carbohydrates
through oxidation (that is, catabolic pathways)
Example:
15 mg of triacylglycerols
= ~1,350 mg of glycogen
Another Lipid Class: Waxes
Esters of
Long chain (14-36 carbon) saturated/unsaturated fatty acids
+ Long chain (16-30 carbon) alcohols
Melting points are higher than triacylglycerols
- Waterproofers for feathers
- Prevent evaporation from holly, rhododendron, poison ivy
- Polishes and lotions
- Derived from palm, lanolin (sheeps wool), beeswax
88 carbons
Major component of beeswax
Three General Classes of Membrane Lipids:
Organized by Backbones
Glycerophospholipids
Spingolipids
Sterols
Two fatty acids joined to glycerol
One fatty acid + one fatty amine (sphingosine)
Rigid core of four fused hydrocarbon rings
Above three are membrane lipid classes
Phospholipids
Some glycerophospholipids and sphingolipids with
a polar head group + fatty acid
Glycolipids
Some sphingolipids with a simple sugar
instead of phosphate
*There are many variations on these themes
What is the backbone?
What is / where are the R -groups?
Note that this is only part of the figure from the book. You are responsible for the entire figure
Glycerophospholipids (Phosphoglycerides):
Derivatives of Phosphatidic Acid
1. General Structure
Glycerol: C1 fatty acid
C2 fatty acid
C3 phosphate + polar head group (alcohol)
Glycerol is not chiral
BUT
Glycerol + phosphate is chiral
.
Phosphodiester bond
for head group
*No known biological significance to the different fatty acids and head groups
Parent Molecule
Ring form with
2 phosphates
Ether Lipids
Phospholipids with fatty acids in ether linkages
About 50% of heart phospholipids are Plasmalogens
Functional significance of ether, rather than ester, linkage not known
MAY function in membrane trafficking, anti-tumor agents
- No strong evidence supports positive roles in health
2. Sphingolipids
Structure
- Derived from Sphingosine
3-carbon backbone
- Sphingosine + long chain FA + Polar Head Group
Head group joined by phosphodiester or glycosidic
linkage
Ceramide
FA is attached to an amide at C2 =
The Parent sphingolipid
Function
- Cell recognition molecules, especially in neurons
- Blood group determinants
(A, B, O System)
C2 amide
Parent Molecule
2. Sphingolipids
Three Classes of Sphingolipids: differ in head groups
Class 1. Sphingomyelins
Head group = phosphocholine or phosphoethanolamine
Therefore, they are phospholipids
Similar to phosphatidylcholine:
No net charge on the head group
at physiological pH
Three Classes of Sphingolipids
Class 2. Glycosphingolipids
Mostly on the outer face of the plasma membrane
Head groups have one or more sugars on C1 of ceramide
Cerebrosides: one sugar on ceramide
Globosides: more than one sugar on ceramide
No net charge at pH 7.0
Neutral Glycolipids
2. Sphingolipids: Clinical / Forensic Significance
Glycosphingolipids
Blood Group Antigens
The differences are in the
carbohydrates
Sphingolipids and Disease
Three Classes of Sphingolipids
Class 3. Gangliosides
- Polar head groups = Oligosaccharides
- Carbohydrate portion has one or more
N-acetylneuraminic acids (a sialic acid)
Another Lipid Class: Galactolipids
No nitrogen:
Different from Glycosphingolipids
No phosphate: Plants conserve phosphate
Located in the thylakoid membranes of chloroplasts
- The most abundant membrane lipids on earth
1,2-diacylglycerol + 1 or 2 galactose at C3
Metabolism of Phospholipids and Sphingolipids
Signaling products result from cleavage
of certain lipid bonds by specific enzymes
Membrane lipids become signaling molecules
Phospholipases cleave phospholipids
Product is a Lysophospholipid
Functions in signaling
One secondary product is a fatty acid
Metabolism of Phospholipids and Sphingolipids
Different signaling products
result from cleavage
of certain bonds by
specific enzymes
- Diacylglycerol
Fatty acid product
Fatty acid product
1,2-diacylglycerol +
Phosphorylated Inositol
products
Sterol Structure: Four Fused Carbon Rings
Sterols are not synthesized by bacteria
Sterol Nucleus
- Three 6-carbon rings
- One 5-carbon ring
- Nearly planar
Produced from simple
precursor: acetate
CH3-COOTwo carbons
Functions of Sterols
Passive Role:
Structural lipids
- In nearly all cells but bacteria
- Cholesterol is amphipathic
nonpolar tail
polar head group +
- Alters membrane fluidity and assists in lipid transport
At low temp: prevents hydrocarbon crystalization
At high temp: decreases overall fluidity
Active Roles:
Diverse
- Steroid hormones are potent signals for gene expression
- Bile acids (derived from cholesterol) emulsify fats to make
them more accessible for degradation
- Precursors for vitamins and hormones
LIPIDS AS SIGNALS, COFACTORS AND PIGMENTS
Phosphatidylinositols: intracellular signals and cofactors
Function: Second Messengers
A signal induces the activation of reactions that produce
other molecules and amplify the initial signal
- The other molecules are second messengers
- The amplification may be in several steps and is
part of a Signal Transduction Pathway
Phosphatidylinositol resides in the plasma membrane
- Can become phosphorylated in several places
- Derivatives are Second Messengers
Phosphatidylinositols
Phosphatidylinositol 4,5-bisphosphate (PIP2)
A lipid on the inner surface of plasma membrane
Functions to
- Bind cytoskeletal proteins
- Bind specific soluble proteins
to promote membrane fusion
- Bind second messengers
after receptor-ligand interactions
PIP2
Phospholipase C
inositol 1,4,5-trisphosphate (IP3) + diacylglycerol
IP3 is a second messenger
- Causes release of
intracellular calcium
- Calcium is a
second messenger
and cofactor for
Protein Kinase C
Diacylglycerol is a
second messenger and
cofactor for Protein Kinase C
Protein kinase C becomes
active
- Phosphorylates proteins
Part of One Signaling Pathway
Eicosanoids: Intercellular Messengers
Paracrine Hormones: act near where they are released
Diverse Functions:
Inflammation
Pain response
Blood clot formation
Fever
Regulation of blood pressure
Gastric acid release
Derivatives of Arachidonic acid (20 carbons)
Three defined classes
Class 1. Prostaglandins
One 5-carbon ring
Two groups with many subtypes
PG E = ether-soluble
PG F = phosphate (water) soluble
Regulate cAMP synthesis through Adenylate Cyclase
cAMP is a Second Messenger and a co-factor
Eicosanoids
Class 2. Thromboxanes
Eicosanoids
One 6-carbon ring
Produced by platelets
Involved in platelet-derived activities like clotting
Synthesis is inhibited by non-steroidal anti-inflammatory drugs
Ibuprophen, aspirin, acetaminophen, etc
Class 3. Leukotrienes
Contain 3 conjugated double bonds
Open (no cyclic) structure
Serve as signaling molecules
Overproduction causes asthma attacks
and contributes to anaphyllactic shock
Eicosanoids
Steroid Hormones: Messengers Between Tissues
Oxidized derivatives of sterol
- Sterol nucleus but no alkyl chain
Move through the blood stream
as a complex with proteins
- Bind to specific receptors
- Bind with high affinity:
Low concentrations
have maximal effects
- Induce gene expression and
changes in metabolism
Functions:
- Gender hormones
- Anti-inflammatory drugs
- Treat asthma and arthritis
Vitamins A and D: Example of
Lipids That are Hormone Precursors
EXAMPLE:
Vitamin A (Retinol)
Derivatives are metabolic and cell regulators
Retinoic acid regulates epithelial cell gene expression
through nuclear receptors
Retinal: a photosensitive pigment in rod and cone cells
- Produces a neuronal response
- Precursor is -carotene
Deficiencies lead to skin and eyes disorders
(wrinkles and night blindness)
Vitamin A: Essential for Sight
Ring + Hydrocarbon
Dietary Vitamin Biologically active
LABORATORY ANALYSIS OF LIPIDS
Lipids are unlike proteins in physical properties
They are studied in organic, non-polar solvents
- Not aqueous solutions
In general, separation of mixtures of lipids utilizes:
- Differences in polarity
- Differences in solubility in non-polar solvents
Lipid Extraction in Organic Solvents
Neutral lipids
Ethyl ether, chloroform or benzene
Membrane Lipids Polar Organic Solvents
(ethanol or methanol)
Solvents reduce weak interactions
i.e. Changing bonds changes structure
Method
Phase extraction using chloroform : methanol : water
1
:
2
: 0.8
This is one phase and extracts all lipids
Add more water and it separates into two phases
Water/methanol phase on top with proteins and sugars
Organic phase on the bottom with lipids
Adsorption Chromatography:
Separation of Lipids Based on Polarity
Column Chromatography
1. Column is packed with insoluble polar material
(silica)
2. Sample is added
Polar Lipids bind to the column
Neutral Lipids do not bind to column
3. Column washed with solvents of increasing polarity
Neutral lipids are eluted in the first wash
(chloroform)
Other lipids are eluted using solvents of increasing polarity
Very polar or charges lipids are eluted with alcohols
Adsorption Chromatography:
Separation of Lipids Based on Polarity
Thin-layer Chromatography
Same principle as column chromatography, except:
A plate of glass is coated with silica
Sample is applied as a spot or line to the bottom of the plate
Bottom of plate (up to the spot) is placed into solvent
The solvent rises into the silica through capillary action
- Lipids separate
The lower the lipid affinity,
the more the lipid moves
Lipids are detected with
iodine vapors, fluorescence
or specific dyes
Two-dimensional Thin-layer
Chromatography
Two-dimensional Thin-layer
Chromatography
Gas-Liquid
Chromatography
Separates charged
lipids best
Separates lipids based on:
- Solubility
- Volatility in solvent
Goes from liquid to gas states with increased temp.
Many lipids are naturally volatile
Those that are not volatile must be derivatized
in order to separate them
Method
1. Add sample into column packed with inert material
2. Heat the column to volatilize the lipids to be separated
3. Force the volatilized lipids through the column using an inert gas
4. Order of elution depends on
- Nature of the solid column support
- Boiling point of the lipid (or derivative)
