General Biochemistry II

MOLB4610/5610
Lecture 10
Biosynthesis of membrane
phospholipids
Lehninger Ch. 21 pg. 791 - 799
Todays lecture
Membrane behavior
Distribution of phospholipids
Phospholipid synthesis in bacteria
Phospholipid synthesis in eukaryotes
Plasmalogen synthesis
Lipid trafficking
Membrane phospholipids
PS Phosphatidylserine
PE Phosphatidylethanolamine
PC Phosphatidylcholine (= lecithin)
PG Phosphatidylglycerol
PI Phosphatidylinositol
DPG Diphosphatidylglycerol (=Cardiolipin)
Why are there so many
different phospholipids ?
Membranes contain mainly proteins and lipids
Phospholipid composition dictates membrane
behavior
Membranes requiring movement within membrane
More unsaturated fatty acids (PE and PS often have
higher amounts of polyunsaturated fatty acids)
Lower packing densities (kinks decrease density)
Shorter fatty acid tails - fewer hydrophobic interactions
Higher membrane fluidity
Cell compartment specific membrane composition
cardiolipin found exclusively at the inner mitochondrial
membrane - decreased leakage
Distribution of membrane lipids
Animals:
major components are PE, PS, PC (= lecithin), &
cholesterol
cell membranes of the central nervous system
contain additional lipids such as sphingomyelin,
myelin, cerebrosides, gangliosides
Plants
PE and PC predominate
PI and PG are present as well
cholesterol is absent but replaced by phytosterols
Bacteria
major components are PE and PG
PC is rarely present, sterols are absent
Phospholipid synthesis in E. coli
Requires an activated component


Both phosphate groups in PP
i
come from CTP
CDP-diacylglycerol can be converted to:
a) Phosphatidylserine (PS) and subsequently to PE
b) Phosphatidylglycerol-3-phosphate (PG)
Phosphatidate + CTP CDP-diacyglycerol + PP
i

pyrophosphate
linkage
Phosphatidate
Synthesis of phosphatidylserine (PS)
& phosphatidyl- ethanolamine (PE)
1) Phosphatidylserine synthase reaction


2) Phosphatidylserine decarboxylase reaction
CDP-diacyglycerol + serine Phosphatidylserine

PS synthase
Phosphatidylserine Phosphatidylethanolamine

PS decarboxylase
CO
2

CMP
H
+

Lehninger pg. 793
Synthesis of
phosphatidylglycerol (PG)
1) Phosphatidylglycerol 3-phosphate synthase


2) Phosphatidylglycerol 3-P phosphatase
CDP-diacyglycerol + glycerol-3-P Phosphatidylglycerol-3-P

PG-3-P synthase
Phosphatidylglycerol-3-P Phosphatidylglycerol
PG-3P phosphatase Pi
CMP
H
2
O
Synthesis of cardiolipin
Cardiolipin synthase reaction
(Bacteria vs. eukaryotes)
BACTERIA

2 PGs glycerol + cardiolipin
EUCARYOTES

CDP-diacyglycerol + PG*
CMP + cardiolipin

* Phosphatidylglycerol is made
exactly as in bacteria
EUCARYOTES
CDP-diacylglycerol + inositol
CMP + Phosphatidylinositol

PI and phosphorylated PI derivatives
play an important role in signal
transduction
Eucaryotic synthesis of
phosphatidylinositol (PI)
PI kinase
Eucaryotic PS synthesis
In yeast PS is synthesized like in bacteria from CDP-
diacylglycerol
In mammals PS is NOT derived CDP-diacylglycerol !
Instead it is derived from PE via a displacement of
headgroups (Serine - ethanolamine)
Q: Where does PE come from in mammals?
PS
PE
IN YEAST
A
A
PE synthesis Mammals vs. Bacteria
1) Ethanolamine Phosphorylethanolamine
ATP ADP
2) Phosphorylethanolamine CDP-ethanolamine
CTP PP
i

3) CDP-ethanolamine Phosphatidylethanolamine
Diacylglycerol CMP
Phosphatidylserine Phosphatidylethanolamine

PS decarboxylase
CO
2

MAMMALS
EUCARYOTES & BACTERIA
PC synthesis in yeast
The amino group of the phosphoglyceride
is methylated 3 times using S-adenosyl-
methionine as the methyl donor.

PE
This reaction also takes place in
bacteria (Stryer pg 718) and in
the liver of mammalian cells.
Although yeast is a eucaryote it
shares share several reactions
with bacteria.
PC synthesis
in mammals
1) Choline kinase mediated
phosphorylation of choline

2) Activation of phosphocholine by
CTP via CTP -choline cytidylyl
transferase

3) Replacement of CMP by
diacyglycerol resulting in PC.

This mechanism used in most
mammalian cells.
Plasmalogen
Ether lipid
Ether: R1-O-R2
Ether-linked alkene: CH
2
-O-CH=CH-(CH
2
)
n
-CH
3

~ 1/2 of heart phospholipids
produced mainly in peroxisomes
Other ether lipids (platelet-activating factor)
CH
2
-O-CH=CH-(CH
2
)
n
-CH
3

CH-O-CO-(CH
2
)
n
-CH
3
CH
2
-O-PO
3
-CH
2
-CH
2
-N(CH
3
)
3
(+) (-)
Plasmalogen synthesis
Step-by-step summary of reactions
backbone = DHAP
Acylation of C
1
in DHAP by fatty acyl CoA
Exchange of an alcohol saturated fatty alcohol
for the carboxylic group at C
1

Reduction by NADPH
Acylation by a second acyl CoA at C
2

Headgroup (ethanolamine) attachment at C
3

Oxidation to alkene via mixed function oxidase
Plasmalogen
synthesis
Platelet activating factor (PAF)
Ether lipid
Acetyl group at C
2
of DHAP backbone
instead of fatty acid
Synthesis similar to plasmalogen
from DHAP backbone
subnanomolar concentration of PAF
induce aggregation of blood platelets
induce muscle contractions
activate the immune system
also the mediator of anaphylactic shocks
Sphingolipid synthesis
Four stages:
1. Synthesis of sphinganine
Requires palmitoyl-CoA + serine (backbone)
Reduction of keto group with NADPH
2. Attachment of a fatty acid to form N-acylsphinganine
3. Desaturation of sphinganine to yield N-acylsphingosine
4. Attachment of the headgroup to produce a sphingolipid
a) Cerebroside (head group e.g. Glucose)
Glycosidic linkage instead of phosphodiester bonds
b) Sphingomyelin (head group e.g. choline)
Lipid trafficking
Lipids that are synthesized at smooth ER
need to relocate to target organelle
Free diffusion is unlikely due to hydrophobic
nature of lipids

Specific transport to target compartments
Requires vesicles that bud from the Golgi
complex