BIOSYNTHESIS

OF
MEMBRANE
LIPIDS
.Floro B. Madarcos, MD
Dept. Of Biochemistry
AMA School of Medicine
 CLINICAL SCENARIO

J. T., a 2-day old premature (33 wks.), born by cesarian section

to a 28-year old primipara, with a birth of 1.95 kgs, developed difficulty
,of breathing 30 minutes after birth manifested as tachypnea, cyanosis
.grunting, subcostal and intercostal retractions
He was placed inside the incubator, given fluids and hooked to a
.respirator
He was given a synthetic lung surfactant. Dramatic improvement
.was noted
 OBJECTIVES
At the end of the lecture, the student should be
able to discuss membrane lipids in terms of:
• Review of the basic composition and the fluid
mosaic model of biologic membranes
• Chemistry – general structure, types and
characteristics
• Synthesis
– Phospholipids, sphingolipids and glycosphingolipids
• Intracellular sites, precursors, mechanisms, enzymes
• Interconversions – base-exchange, methylation,
decarboxylation
– Ether phospholipids – precursors, enzymes
• Deficiency syndromes
• Degradation
• Functions
 I. REVIEW OF BASIC COMPOSITION
BIOLOGIC/CELL MEMBRANES
OF BIOLOGIC/CELL MEMBRANES
– Proteins - 50%
– CHOs - 5%
• Glycoproteins
• Glycolipids
– Lipids - 45%
• Phospholipids
• Sphingolipids
– Sphingomyelin
• Glycosphingolipids
– Cerebrosides
• Cholesterol
II. REVIEW OF THE FLUID MOSAIC MODEL
OF BIOLOGICAL MEMBRANES

Cholesterol

Peripheral
or extrinsic

Peripheral
Integral or extrinsic Integral
or intrinsic or intrinsic
III. TYPES OF MEMBRANE PHOSPHOLIPIDS
(GLYCEROPHOSPHOLIPIDS OR
PHOSPHOGLYCERIDES)

• Phosphatidylethanolamine (Cephalin)
• Phosphatidylcholine (Lecithin)
• Phosphatidylserine
• Phosphatidylinositol
• Phosphatidylglycerol
• Cardiolipin(Diphosphatidylglycerol)
 IV. GENERAL STRUCTURE OF A PHOSPHOLIPID
(GLYCEROPHOSPHOLIPID or PHOSPHOLYCERIDE)
O
Fatty acid Fatty acid
).unsat(
II ).sat(
O 1CH2 – O – C – C – R1
II I
.Organic molec
R2 - C – O - 2C – H O Nitrogenous
I II polar or hyro-
philic grp.
Glycerol 3CH2 – O – P – O - X
backbone
I
O- Phosphodiester
bridge

d i a c y l g l y c e r o l - 1,2
OH OH
CH3
Polar Head Groups O-
I
-CH2-CH2-N-CH3 CH2-CH-NH3- CH2-CH-CH2OH-
HO
I I CH2-CH2-NH3- I
OH OH
CH3 -
COO
OH
Choline Serine Ethanolamine Glycerol
Inositol
V. ANOTHER REPRESENTATION OF PHOSPHOLIPID STRUCTURE
X = Variable polar group
e.g. Choline
Ethanolamine
Serine
X Inositol
O Glycerol
POLAR

O–P=O
- Phos-
phate
H

POLAR
H2C – C – CH2
1 2 3
O O Glycerol
O=C C=O

FA tail
Double bond
NONPOLAR

(kink in tail)

NONPOLAR
Unsaturated FA
Bilayer
-Oleic
-Linoleic
-Linolenic
Saturated FA -ARA, EPA, DHA
-Myristic FA tail
-Palmitic FA tail length
-Stearic (16-20 C atoms)
 VI. STRUCTURES OF SOME PHOSPHOLIPIDS
Longer chain PUFA O Saturated FA
Linoleic acid (18:2)
II Palmitic acid (16:0)
Arachidonic acid (20:4)
Docosahexaenoic acid (22:6) O 1CH2 – O – C – C – R1 Stearic acid (18:0)
II I
R2 - C – O - 2C – H O
I II +
3CH2 – O – P – O – CH2-CH2-NH3
I
O-
Phosphatidylethanolamine (PE; Cephalin; neutral; 2nd most
Longer chain PUFA common =20-30%; inner leaflet)
Linolenic acid (18:3) O Saturated FA
Linoleic acid (18:2) Palmitic acid (16:0)
Oleic acid (18:1
II
Stearic acid (18:0)
Arachidonic acid (20:4) O 1CH2 – O – C – C – R1

II I
R2 - C – O - 2C – H O
I II +
3CH2 – O – P – O – CH2-CH2-N-(CH3)3
I
O-
Phosphatidylcholine (PC; Lecithin; neutral; most abundant = 50%; outer leaflet)
 VII. STRUCTURES OF SOME PHOSPHOLIPIDS
O
II
O 1CH2 – O – C – C – R1

II I
R2 - C – O - 2C – H O
I II +
3CH2 – O – P – O – CH2-CH-NH3
I I
O- COO-
Phosphatidylserine (PS; acidic; 5-10%; inner leaflet)
Arachidonic
O
acid (20:4) Stearic acid (18:0)
(almost exclusively)
II
(almost exclusively)
O 1CH2 – O – C – C – R1

II I
R2 - C – O - 2C – H O
I II
O H
3CH2 – O – P O
I HO 1 6 O P – OII
DAG + IP3
-

O- OH
OH
5 H I -

2 O
-

H 3 4 H
H O
Phosphatidylinositol (PI; acidic)
-O- P-O
(Phosphatidylinositol 4,5-bisphosphate) I
-O
 VII. STRUCTURES OF SOME PHOSPHOLIPIDS
O
II
O 1CH2 – O – C – C – R1

II I
R2 - C – O - 2C – H O
I II
Phosphatidic 3CH2 – O – P – O – CH2-CH- CH2OH
acid I Phosphatidic
I OH acid
O-
Phosphatidylglycerol (PG; acidic; primariy in
mitochondrial membranes & pulmonary surfactant )
O
II O
O II
1CH2 – O–C–C–R1
CH2 – O – P – O – 1CH2 O
II I I
I I II
R2 - C – O - 2C – H O HO–C – H O -
2CH – O - C – R3
I II I
I
3CH2 – O – P – O –
CH2
3CH2 – O – C – R4
I Glycerol II
O backbone
-
O
Cardiolipin (Diphosphatidylglycerol; very acidic; primarily in mito.
membranes of metabolically active tissues, e.g. heart)
 VIII. STRUCTURE OF ETHER
GLYCEROLIPIDS
R1 (palmitate
Vinyl ether linkage
)C16

H H Ethanolamine
O 1 CH 2 – O – C = C – (CH )
2 15 - CH 3 or
II I Choline
R 2 - C – O - 2C O
–H
I II +
3CH2 – O – P – O – CH2-CH2-NH3
I
O-
Plasmalogen
(Ethanolamine plasmalogen - ↑ in myelin, ↓ in heart) )
(Choline plasmalogen - ↓ in myelin, ↑ in heart)
Glycerol IX. OVERVIEW OF DE NOVO GLYCEROPHOSPHOLIPID SYNTHESIS
Glycerol kinase (SER, outer mito. membrane)
Liver activated 2
fatty acids CoA
Glycerol 3-PO4
Phosphatidic Acid
Dehydrogenase )Phosphatidate; 1,2-diacylglycerol PO4(
GPAT
Adipose CTP
tissues Phosphatidic acid CDP-Diacylglycerol
Liver
DHAP phosphatase (PAP) synthase (CDS)
or lipin
mechs 2
Choline
PPi Inositol
Head Pi A B Serine
Ethanolamine
group Glycerol 3-PO4
CTP Diacylglycerol 1,2 CDP-diacylglycerol
)DAG( )CDP-DAG( Head group
CDP- head
group TAGs

CMP CMP

Glycerophospholipid Glycerophospholipid
Phosphatidylcholine AdoMet 3
Phosphatidylinositol
CO2
Phosphatidylethanolamine Phosphatidylserine
”be“ Phosphatidylglycerol
Phosphatidylserine ”be“ moles 2
Cardiolipin
 H O
|
X. SYNTHESIS OF PHOSPHATIDATE ||
H – C1 – OH CH3 – (CH2)x – C
De novo
| synthesis |
H – C2 = O
Gl
yc Fatty acid* O
-P rale H
| O de I
,ad ro eh hyd
d 4 d Thiokinase
H – C3 – O – P ipo gen y-3 e H – C1 – OH
se as ATP CoA
| ) l i v ti s s e |
er ue H – C2 – OH
H s( AMP
| PPi + O
Dihydroacetone ||
PO4 (DHAP ) H – C – O – P
3 CH3 – (CH2)x – C – SCoA
NADH |
H
Fatty acyl CoA 2
+
H+ +
NAD
Glycerol 3-PO4* Gl CoA
yc
Ac e
ylt rol 3
Glycolysis ran -PO
Glycerol kinase ) s
GP fer 4 O
H r
)liver( ate- AT( se a II
| lim
H – C1 –O H ADP itin O C1H2O-C-R1
Pi + g II I
|
H – C – OH R2-C-O-C2
| ATP I
H – C3 – O – H
+ CoA-C–SH 2
3H2 – O-P3
2

| Phosphatidate
H (1,2-diacyl-
Glycerol building blocks in phospholipid synthesis*
TAG glycerol PO4)
XI. SYNTHESIS OF PHOSPHATIDYLCHOLINE & PHOSPHATIDYLETHANOLAMINE
O Sat. FA +
)ER/outer mitochondrial membrane( OH-CH -CH -NH II 2 2 3

O C1H2-O- C-R1 Ethanolamine

PUFA (ex. oleic acid, 18:1) ATP
II I Ethanolamine
R2-C -O-C2-H kinase (cyto) ADP rate(
I Phosphoethanolamine )limiting
+ CTP Phosphoethanolamine
OH-CH2-CH2-N(CH3)3 C3H2-OP32- Cytidyltransferase
Phosphatidate (ER)
PPi (cyto/ER)
Choline Fatty (1,2-diacylglycerol PO4) +
acyl CoA CDP -O-(CH ) -NH
CoA
2 2 3
PAP Pi CDP-Ethanolamine
ATP Choline
*1,2-Diacyl-
kinase
TAG Phosphoethanolamine
ADP (cyto.)
CTP PPi
acyltrans- Glycerol DAG transferase (ER)
ferase (DAG)
+ CMP
Phospho- CDP-O-(CH2)2-N(CH3)3 0
choline Phosphocholine
Cytidyltransferase CDP-Choline II
(PCT; cyto/ER) ) Phosphocholine O C1H2-O- C-R1
(rate-limiting)
1 DAG transferase
(ER) II I
CMP R2-C -O-C2H O
Lungs
0 I II +
*branch pt. in II C3H2-O- P-O-CH2-CH2-NH3
s
TAG & O C1H2-O- C-(CH2)14-CH3 ion I
phospolipid t
a PEMT 3 SAM O- Phosphatidyl-
II I l
synthesis
CH3-(CH2)14-C-O-C2H t hy )liver( 2 ethanolamine
O e
PAP = Phosphatidic acid II M 3 SAH
I 3 +
phosphatase C3H2-O- P-O-CH2-CH2-N-CH3-CH3-CH3
PEMT = phosphatidylethanolamine Phosphatidylcholine
methyl transferase
I
O-
 STRUCTURE OF DIPALMITOYL
PHOSPHATDYLCHOLINE (DPPC)

O
II
H2C1 – 0 – C-(CH2)14 –CH3
O Palmitic acids
II
Glycerol H2C2 – 0 – C-(CH2)14 –CH3
backbone O
I CH3
H2C3 – 0 – P- CH2 –CH2 – N CH3
II
CH3
O

Choline
 XII. ACUTE RESPIRATORY DISTRESS SYNDROME
(ARDS) AND SYNTHETIC SURFACTANT
1. Chest radiographs of a
premature infant with
acute respiratory distress
syndrome before and after
surfactant treatment.

2. Left: Initial radiograph

shows poor lung
expansion, air broncho-
gram, and reticular
granular appearance.

3. Right: Repeat chest

radiograph obtained when
the neonate is aged 3
hours and after surfactant
therapy demonstrates
marked improvement.
 XIII. INTERCONVERSIONS OF PHOSPHATIDYLETHANOLAMINE
AND PHOSPHATIDYLSERINE
0
II
O 1CH2-O-C-R1
II I
R2-C -O-2CH O
I II +
3CH 2
P-
-O- O-CH2-CH2-NH3
+NH3 I
O-
I
HO-CH2-CH-COO-
*Phosphatidylethanolamine(PE)
Serine CO2
Phosphatidylethanola- Base“ Minor
mine: serine transferase Phosphatidylserine
”exchange pathway decarboxylase
(Phosphatidyl serine ).liver mito(
synthase 2; ER) )major route( (mitochondrial)
O +
H
II
HO-CH2-CH2-NH3+ O 1CH2-O-C-R1
II I
Ethanolamine R2-C-O-2CH O +NH3
ways for PE 3* I II I
3CH2-O- P-O CH2-CH-COO
-
synthesis
Phosphatidylserine
 XIV. INTERCONVERSION OF PHOSPHATIDYLCHOLINE
AND PHOSPHATIDYLSERINE
0
II
O C1H2-O- C-(CH2)14-CH3
II I
CH3-(CH2)14-C-O-C2H O
I II +
C3H2-O- P-O-CH2-CH2-N-CH3-CH3-CH3
+NH3
I Phosphatidylcholine (lecithin)
I
O-
HO-CH2-CH-COO -

Serine Phosphatidylcholine
Base“
serine transferase
”exchange (Phosphatidylserine
synthase I; ER)
+
OH-CH2-CH2-N-CH3-CH3-CH3
O
Choline
II
O 1CH2-O-C-R1
II I
R2-C-O-2CH O +NH3
I II I
3CH2-O- P-O CH2-CH-COO
-

Phosphatidylserine
 XV. SYNTHESIS OF PHOSPHATIDYLSERINE & PHOSPHATIDYLINOSITOL
O 2 Pi O Phosphatidate
II II
O 1CH2-O-C-R1 CTP PPi O C-R1 NH2
1CH2-O- Cytidine
II I II I monoPO4
R2-C-O-2CH O R2-C-O-2CH O O N
I II CDP phospha- I II II
3CH2-O-
P-O -
tidate 3CH2-O-
P-O- P-O CH2 O
N
I cytidyltrans- I I O
O - ferase O- O-
H H
Phosphatidate +NH3 CDP-Diacylglycerol
(1,2-diacylglycerol I H H
OH OH
PO4) HO-CH2-CH-COO- Inositol
Serine Phosphatidylinositol
Phosphatidylserine synthase
synthase Some Bacteria & CMP
bacteria eukaryotes
CMP O
II
O O CH2-O- C-R1 (Stearic acid)
II II I
O CH2-O- C-R 1 (Arachidonic R2-C-O-CH O
II I acid) I II
R2-C -O-CH CH2-O- P O H
O +NH3 O PO32-
I II I 0H 1 6
I O - OH
CH2-O- P-O CH -CH-COO- 2 5
ways 2*
2 Prostaglandins OH H
I H 3 4
for PS H
O- H
O
synthesis
Phosphatidylserine* PO32-
Phosphatidylinositol 4,5-bisP
 XVI. CONVERSION OF PHOSPHATIDYLINOSITOL TO PIP2 AND IP3
O O Site of cleavage by
II II Phospholipase C
O CH2-O- C-R1 O CH2-O- C-R1
II I II I
R2-C-O-CH O 2ADP R2-C-O-CH O
H OH
2 ATP + 2Pi H OPO3-
I II I II
CH2-O- P O H CH2-O- P O 5 H
I I
OH OH
O- 1 OH OH 4 O- 1 OH OH 4
H OPO3- Kinase H OPO3-
H H H H
Phosphatidylinositol Phosphatidylinositol 4,5-bisphosphate (PIP2)

Phospholipase C
O H2O
II
R1-C-O-CH2 O
O
+ II H OPO3-
II -
O P O 5 H
R2-C-O-CH2 I
OH
I O- 1 OH OH 4
CH2-OH
2nd messengers H OPO3-
Diacylgycerol (DAG) H H
Inositol 1,4,5-trisphosphate (IP3)
Phosphatidate
CTP
PPi XVII. SUMMARY OF INTERCONVERSIONS OF
CDP-diacylglycerol
CHOLINE, ETHANOLAMINE AND SERINE 0
Serine
O +
H II
II Phosphatidylserine CO 2 O 1CH2-O-C-R1
O 1CH2-O-C-R1 decarboxylase II I
II I R2-C -O-2CH O
R2-C-O-2CH O +NH3 I II +
I II I 3CH 2
P-
-O- O-CH2-CH2-NH3
3CH2-O- P- O CH2-CH-COO- I
O-
Phosphatidylserine
Phosphatidylethanolamine

Ethanolamine Serine
Phosphatidylcholine Phosphatidylethanola 3
Choline serine transferase mine AdoMet
0 methyltransferase
II (PEMT; liver ) 3
O C1H2-O- C-(CH2)14-CH3 AdoHcy
Serine II I
CH3-(CH2)14-C-O-C2H O
I II +
C3H2-O- P-O-CH 2-CH 2-N-CH 3-CH3-CH3
I Phosphatidylcholine
O-
XVIII. SYNTHESIS OF CARDIOLIPIN
Glycerol
ATP
Glycerol kinase
ADP + Pi

Glycerol 3-PO4
Fatty acyl CoA
CoA
Acyltransferase
Fatty acyl CoA
CoA
Phosphatidate
CTP

PPi
CDP-diacylglycerol
Glycerol 3-PO4
Synthase
CMP
Phosphatidylglycerol 3-PO4
H2O
Phosphatase
Pi
Phosphatidylglycerol (PG)
Cardiolipin synthase PG
(eukaryotes) Glycerol

Diphosphatidylglycerol (CARDIOLIPIN)
 H2C1-OH XIX. SYNTHESIS OF ETHER
I
C2=O GLYCEROLIPIDS - PLASMALOGEN
O I O
II H2C3-O-P II
R-C-SCoA DHAP R1-CH2-CH2-C-SCoA
Fatty acyl NADPH 2
Acyl transferase
CoA
Peroxisomes Platelet Activating
O
CoA II NADPH Factor
C1H2-O-C-R R1-CH2-CH2-OH +
H+ )PAF(
I Long chain( +NADP
C2-O O )FA alcohol Reductase
II Acyltransferase
II
O-P-O- Phosphohydrolase
C3H2
I Synthase Acyl CoA H2O Pi
acylDHAP-1 O-
Long chain alcohol
O C1H2-OH-CH2-CH
O 2-R1
C1H2-OH-CH2-CH2-R1
II II I
I Reduction of C2 to an R -C-O-
R-C-O- C - H
C2- O O alcohol, addition of a
2 2

II I
Unsat. I fatty acid and
O-P-O - C3H2- OH
C H
alkenyl 3 2 dephosphorylation
Ether group I alkyl-2-acylglycerol-1
linkage O- CDP-Ethanolamine
alkylDHAP-1 Transferase
Acyl C1H2-O-CH=CH-R
O 1
CMP
group II I O2 Cyt b5 NADPH
R2-C-O- O Unsat.
C2-H C1H2-O- CH2-CH
O 2-R1
II
I II I
O-P-Ethanolamine
-C3H2 Desaturase R2-C-O-C2-H
O alkyl group
I II
I
alkenyl-2-acylglycerol-1 O- O-P-Ethanolamine
-C3H2
phosphoethanolamine-3 alkyl-2-acylglycerol-1 I
)a plasmalogen; found in myelin sheath( phosphoethanolamine-3 O-
 DHAP
Fatty acyl XX. SYNTHESIS OF ETHER GLYCEROLIPIDS - PLATELET
CoA ACTIVATING FACTOR (PAF)
acylDHAP-1 Glycerol
CDP- backbone
FA alcohol Choline CMP
C1H2-OH-CH2-CHO 2-R1 O 2-R1
C1H2-OH-CH2-CH
II I II I
R2-C-O-
C2=O R2-C-O-
C2=O
alkylDHAP-1
I I
C3H2- OH C3H2- O - P Choline
Transferase 1-alkyl-2-acylglycerol
alkyl-2-acylglycerol-1
3-phosphocholine

Phospholipase A2
Sat. R2-COOH
Acetyl group Ether alkenyl
inkage group (Fatty acyl CoA)
Acetyl
CoA CoA 2-CH2-R1
C1H2-OH-CH
H2-CH2-R O1
II I I
CHO3 - C-
C2- H Sat. HO - C2- H
I I
Choline C3H2- O - P Choline
C3H2- O - P Acyltransferase 1-alkyl-2-lysoglycerol
1-alkyl-2-acetylglycerol 3-phosphocholine
3-phosphocholine (PAF)
 XXI. DEGRADATION OF
PHOSPHOLIPIDS
Lysophospholipid
Free FA +

Phospholipase A1
(mammalian tissues) A1
O
A2 II
Phospholipase A2 C1H2 – O – C – R1 Phosphatidic acid
(mammalian tissues/ O free head group +
pancreatic juice) II
Lysophospholipid
R2 – C – O – C2H D Phospholipase D
O
Free FA +
II
C3H2 – O – P –O - Head group
I
O-

diacylglycerol-1,2
phosphorylated alcohol +
C

Phospholipase C
(liver lysosomes)
 BIOSYNTHESIS OF MEMBRANE LIPIDS:
SUMMARY
• Review:
– Basic membrane composition – CHOs, CHONs, lipids
– Fluid mosaic model – phospholipid bilayer, cholesterol insertion,
random protein distribution, membrane asymmetry & fluidity &
permeability
• Basic phospholipid structure – glycerol backbone,
phosphorylated organic molecules, amphipathic nature - polar &
nonpolar groups
• Biosynthesis – ER, glycerol kinase, glyce. 3-PO4 DH  glycerol
3-PO4  acylations  Phosphatidate; same initial pathway for
TAG synthesis
– 1st mech – DAG: PC “be” PS; PE “sam” PC; PS –CO2 PE; PE “be” PS
– 2nd mech – CDP-DAG: PI; PG, Cardiolipin
• Functions – PI – 2nd messengers (IP3 & DAG) ; DPPC – lung
surfactant; PS – blood coagulation, apoptosis; component of biologic
membranes
• Degradation – phospholipases for hydrolysis of fatty acids,
release of PO4 group and organic head groups.
 REFERENCES
• Basic Medical Biochemistry: A Clinical Approach, Lieberman,
M., Marks, A.D., 3rd ed., pp. 621-625.
• Harper’s Illustrated Biochemistry , Murray, R.K., et. al., 28th ed.,
pp. 205-211.
• Textbook of Biochemistry with Clinical Correlations , Devlin,
T.M., 7th ed., pp. 708-718.
• Biochemistry, Lippincott’s Illustrated Reviews, Champe, P.C.,
Harvey, R.A., Ferrier, D.R., 4 th ed., pp.201-208.
• Biochemistry, Campbell & Farrell, 6th ed., pp. 628-630.
• Biochemistry, Mathews, C.K., Van Holde, K.E., Appling, D.
R. Anthony-Cahill,S.J., 4th ed, pp. 364-369, 781-790.
• Medical Biochemistry, Baynes, J.W., Dominiczak, M.H., 4 th ed.,
p. 26, pp. 369-378.