Respiratory Module

biochemistry
for PC-I student
Surfactant metabolism

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Surfactant
• Lung surfactant is synthesized in the endoplasmic reticulum of the
alveolar type II epithelial cell.
• Stored in lamellar bodies, an intracellular storage form.
• Lung surfactant is secreted via exocytosis from type II cells involving
fusion of lamellar bodies with the plasma membrane in response to
extracellular signals.
• Once secreted from type II cells, the extracellular pool of phospholipid
within lamellar bodies transforms into a surfactant film that lines the
alveolar surface.

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Surfactant
• The composition of surfactant is 90% lipids and 5-10% surfactant
specific proteins.
• The lipid component is made up of phospholipids, triglyceride,
cholesterol and fatty acid.
• The phospholipid composition of surfactant is highly conserved
among mammals.

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Surfactant
• Phospholipid found in surfactant are
Dipalmitoylphosphatidylcholine (lecithin)- accounts 70-80%
Phosphatidylglycerol (PG)- accounts 10%.
 it may play a role in alveolar stability and it regulates the innate immune response.
The remainder are phosphatidylinositol (PI), phosphatidylethanolamine (PE),
and phosphatidylserine (PS).
• Immature surfactant contains higher amounts of PI compared to PG.
• Thus, a low ratio of PG to PI indicates lung immaturity.

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• The protein component of surfactant is lung-specific.
• consists of four proteins designated SP-A, SP-B, SP-C, and SP-D.
• Function of surfactant proteins are
 Structural transformation of lamellar body to tubular myelin (SP-A
and SP-B in the presence of Ca 2+)
 Enhancement of surface-tension lowering properties and promotion
of adsorption of surfactant phospholipids at the air-liquid interface
(SP-B and SP-C).

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 Reuptake by endocytosis of surfactant by type II cells, and the
activation of alveolar macrophages to facilitate surfactant clearance.
• Both SP-A and SP-D possess antimicrobial properties.
• SP-A is chemotactic for macrophages and promotes bacterial
phagocytosis.

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• The primary structure of SP-A is highly conserved among several
species.
• It has two domains;
• the N terminus is collagen-like with Gly-X-Y repeats (where Y is frequently a
prolyl residue), and
• The C terminus has lectin-like properties.
• SP-B and SP-C are highly hydrophobic proteins while SP-A and SP-D
are hydrophilic.
• SP-D is a glycoprotein and has a structure similar to SP-A.

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LAMELLAR BODY (LB) FORMATION
• Pulmonary surfactant is synthesized, packaged, and stored in alveolar
type II epithelial cells as Lamellar body (LB).
• lysosome-related organelles secreted into the alveolar space via exocytosis
that transform into the surfactant film.
• The glycogen stores of fetal type II cells are a site for surfactant PC
synthesis and LB formation.
• Secreted surfactant is internalized by type II cells that can be
incorporated back to LB for recycling or degradation by alveolar
macrophages.

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• LB is characterized by an acidic interior that contains lysosomal
enzymes.
• acid phosphatase and cathepsins C and H
• proteins (CD63/LAMP3, LAMP1).
• These organelles contain concentric and tightly packed lamellae
comprised of DPPC.
• SP-B plays a central role in packaging of surfactant phospholipids into
LB.

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• Both SP-B and SP-C processed in the Golgi are transported to LB
through multivesicular bodies via proteolytic processing.
• In contrast, phospholipids such as PC, DPPC, and PG newly
synthesized in the ER are transported directly to LB.
• These surfactant-associated phospholipids are transported from the
ER to LB via phospholipid transfer proteins and then transverse the LB
membrane.

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• This process can be facilitated by ATP-binding cassette (ABC)
transporters such as ATP-binding cassette A3 (ABCA3).
• ABCA3 is a lipid transport protein that is required for normal synthesis
and storage of pulmonary surfactant in alveolar type II cells.

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• Surfactant biosynthesis is developmentally regulated.
• The capacity for the fetal lung to synthesize surfactant occurs
relatively late in gestation.
• the secretion of surfactant into the amniotic fluid occurs during 30-32
weeks of gestation.

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• The synthesis of surfactants is regulated by glucocorticoids, thyroid
hormones, prolactin, estrogens, androgens, catecholamine, growth
factors, and cytokines.
• Glucocorticoids stimulate lung maturation.
• Thus, glucocorticoid therapy in women in preterm labor prior to 34
weeks of gestation can significantly decrease the incidence of
respiratory distress syndrome in the premature neonates.

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• Thyroid hormones also accelerate fetal lung maturation.
• Insulin delays surfactant synthesis and so fetal hyperinsulinemia in
diabetic mothers may increase the incidence of Respiratory distress
syndrome even in the full-term infant.
• Androgen synthesized in the fetal testis is the probable cause of a
slower onset of surfactant production in male fetuses.
• Estrogen facilitate lung maturation.

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• Prophylactic, or after onset of respiratory distress syndrome,
administration of synthetic or natural pulmonary surfactants
intratracheally to preterm infants improves oxygenation and decrease
pulmonary morbidity.

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Biochemical Determinants of Fetal lung
Maturity
• The biochemical determinants are measured primarily in the amniotic
fluid obtained by amniocentesis.
• In a normal pregnancy, the lung is adequately developed by about the
36th or 37th week.
• Biochemical changes occurring during this period of gestation can be
used to evaluate fetal lung maturity when early delivery is planned.
• Tests used to evaluate fetal lung maturity is:
• lecithin to sphingomyelin (L/S) ratio
• foam stability test (FST), or shake test: measured by its ability to generate stable
foam in the presence of ethanol.
• measurement of lamellar bodies
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• lecithin to sphingomyelin (L/S) ratio in amniotic fluid.
• In a normal pregnancy, the L/S ratio is less than 1 before the 31 st
week, rises to about 2 by the 34th week, to about 4 at the 36th week,
and to about 8 at term (39 weeks).
• The change is due to an increase in lecithin synthesis rather than a
decrease in synthesis of sphingomyelin.

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Biochemical Determinants of Fetal lung
Maturity
• These values vary in normal gestations and in abnormal pregnancies
(due to maternal, fetal, or placental disorders), the ratio may be
elevated or reduced without regard to gestational age.
• A low L/S ratio is not inevitably associated with RDS.
• While an L/S ratio greater than 2 is associated with the absence of
serious RDS, one lower than 2 is not uniformly predictive of the
development of RDS.

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Phospholipids

• Phospholipids can be glycerolipids or sphingolipids.

• Examples of glycerolipids are phosphatidylcholine,
phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol,
and phosphatidylglycerol.

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SYNTHESIS OF PHOSPHOLIPIDS
• All cells, with the possible exception of mature red blood cells, are
capable of synthesizing one or more glycerol phospholipids.
• Most of the reactions involved in phospholipid synthesis occur on the
cytosolic face of the endoplasmic reticulum and Golgi complex.
• The liver is a major site of phospho lipid synthesis
• Two other tissues with a high capacity for phospholipid synthesis are
intestinal enterocytes, type II cells of the lung, which synthesize
pulmonary surfactant

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• Phosphatidylcholines or lecithins
• Phosphatidylcholines are the most abundant phospholipids in animal
tissues.
• Typically contain palmitic, stearic, oleic, linoleic, or arachidonic acid.
• The de novo pathways for phospholipid synthesis use cytidine
triphosphate (CTP) for activation of intermediate species
• The principal pathway of phosphatidylcholine biosynthesis uses
cytidine diphosphate (CDP) choline

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• Many reactions of phospholipid synthesis occur in the endoplasmic
reticulum.
• Choline is first phosphorylated by ATP to phosphocholine, which
reacts with CTP to form CDP choline, from which phosphocholine is
transferred to 2-diacylglycerol.
• The rate-limiting step in this pathway appears to be that catalyzed by
CTP: phosphor choline cytidylyl transferase, which is activated by fatty
acids.

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• Phosphatidylcholine can also be synthesized by the methylation
pathway that converts phosphatidylethanolamine to
phosphatidylcholine, principally in the liver.
• The methyl donor is S-adenosylmethionine.
• Phosphatidylethanolamine-N-methyltransferase transfers three
methyl groups in sequence to produce phosphatidylcholine.

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• Phosphatidylcholine is degraded by phospholipases that cleave
preferentially at specific bonds.
• Choline released is phosphorylated by choline kinase and reutilized in
phosphatidylcholine synthesis.
• However, in liver mitochondria, choline is also oxidized to betaine (N-
trimethylglycine):

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• Betaine functions as a methyl donor (e.g., in methionine biosynthesis
from homocysteine), and it can also be converted to glycine.

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DEGRADATION OF PHOSPHOLIPIDS

Phospholipase A1:-
-found in many mammalian tissues.
-removes fatty acid from C1
Phospholipase A2:-
-found in many tissues and pancreatic juice
-removes F.A at C2
-inhibited by glucocorticoids

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DEGRADATION OF PHOSPHOLIPIDS

Phospholipase C:-
-cleaves phosphate group at C3
-found in liver lysosomes and some bacteria
-role in producing second messengers.
Phospholipase D:-
-found primarily in plant tissues.
-removes the compound with alcohol group on C3

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Reading assignment
• Synthesis of PI, PS, PE and cardiolipin.

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