AROMATIC AMINO ACID METABOLISM IN ANIMALS

Although

animal

cells

do

not

synthesize

aromatic

rings

do

novo,

the

intermediarymetabolism of aromatic amino acids in animals is extensive,and it involves many

important biological processes. These include the biosynthesis of tyrosine from phenylalanine,
the utilization of tyrosine,tryptophan,and histidine in synthesis of biogenic amines,compounds
that serve as hormones and as neurotransmitter. Aromatic amino acid metabolism also provides
the backdrop for much of our early understanding of human biochemical genetics.
Tyrosine Biosynthesis
The only known reaction of aromatic amino acid biosynthesis in animals is the
conversion of phenylalanine to tyrosine, catalyzed by phenylalanine hydroxilase. This interesting
enzyme is a mixed-function oxygenase that uses a pteridine cofactor,tetrahydrobiopterine. The
reaction oxidizes tetrahydrobiopterin to the quinonoid isomer of dihydrobiopterine,as shown in
figure 21.18.

Figure 21.18. The phenylalanine hydroxylase and dihydropteridinereductase reactions.

The

coenzyme

is

regenerated

trough

the

action

of

the

NADPH-requiring

dihydropteridinereductase (analogous,but not identical,todihydrofolatereductase). This enzyme

system occurs almost entirely in the liver.

A hereditary deficiency of phenylalanine hydroxylase is responsible of phenyiketonuria

(PKU),a condition that afflicts about 1 in 10,000 newborn infants in western Europe and the
united States. PKU is an autosomal recessive trait,meaning that two parents heterozygous for the
trait have 1 chance in 4 of having a phenylketonuric child. From the incidence of the disease,we
can estimate that about 2 % of the population are carerrs. In phenylketonuria,phenylalanine
accumulates to very high levels because of the block in conversion to tyrosine,and much of this
phenylalanine is metabolized via pathways that are normally little used-particularly
transamination to phenylpyruvate,and also subsequent conversion of the phenylpyruvate to
phenyllactate. These compounds are excreted in urine in enormous quantities (1 to 2 grams per
day).
If undectected and untreated,PKU leads to profound mental retardation ;the precise
biochemical causation has not yet been identified. Fortunately,PKU can readily be detected at
birth,and many hospitals carry out routine screening of newborns. If the condition is detected
early,the onset of retardation can be prevented by feeding for several years a synthetic diet low in
phenylalanine and rich in tyrosine,to allow normal development of the nervous system. Because
the use of this synthetic diet is quiet expensive,there has been much interest in prenatal diagnosis
of PKU and in identification of heterozygous carriers. The human gene for phenylalanine
hydroxylase has been cloned,providing the basis for a test,using nucleic acid hybridization,that
can be applied an any cell.
In recent years a different form of PKU has been described that results from a hereditary
deficiency of dihydropteridinereductase. This condition is much rarer than classical PKU
and,becausetetrahydrobiopterin is involved in other hydroxylation, much more severe in its
symptoms.
Tyrosine Utilization and Catabolism
In animals, phenylalanine plays no significant role other than as a component of proteins
and a precursor to tyrosine. However, tyrosine plays several important roles in animal
metabolism-as precursor to thyroid hormones, to the biological pigments called melanins, and to
the catecholamines, which serve both as hormones and as neurotransmitter (discussed later in
this capter).
Thyroid hormones stimulate a number of metabolic processes, through activation of the
transcription of particular genes (see Chapter 23). The synthesis of thyroid hormones,principally

thyroxine (T4) andtriiodotothyronine ( T3 ),occurs by a novel pathway,at the level of thyrosine

residues in a specific protein, thyroglobulin. As shown in Figure 21.19,two iodinated thyrosine
residues on the same polypeptide chain react to give a residues of T 3 or T4,which is released by
proteolysis. This protein undergoes degradation to yield the free hormones, which are transported
to their sites of action through the bloodstream. The synthesis of thyroid hormones involves
iodinitation of the thyrosine ring. This process occurs in the thyroid gland, which concentrates
iodide ion from the blood serum for this purpose. One result of iodine deficiency is goiter,a
condition in which the thyroid gland grows abnormally large as it attempts to scavenge all
available iodine. Before iodized salt came into widespread use, goiter was endemic in regions
whose soil was deficient in iodine.

Figure 21.20. Biosynthetic pathways from tyrosine to melanins

The synthesis of melanins (Figure 21.20) occurs in pigment-producing cells, the
melanocytes. For some time it was thought that the entire pathway involves just one enzyme.
However, in mammalian tissues, it is likely that the initial step, the hydroxylation of thyrosine to
3,4-dihydroxyphenylalanine (usually called by the acronym dopa),involves a peroxidase
reaction. Dopa, which also acts as a cofactor for this raction, is the substrate for a subsequent
oxidation to dopaquinone, catalyzed by a copper-containing oxygenase called thyrosinase. The
subsequent reactions, leading to melanins, occur spontaneously in vitro. However, thyrosinase
has been shown to catalyze another reaction, on the branch of the pathway leading to black
melanin-the oxidation of 5,6-dihydroxyndrole to indole-5,6-quinone. This reaction also requires
dopa as a co factor and is inhibited by thyrosine, apparently providing a means for regulation of
pigment formation.
In another branch of the pathway. Dopaquinone reacts with cysteine en route to a related
series of palymers, the red melanins. And individuals pigmentation is determined by the relative
amounts of red and black melanins in the skin. These in turn result from the distribution an
density of melanocytes in the basal layers of the skin, as well as the activities of the pathways
leading to the different melanins. A genetic deficiency of thyrosinasecouse an individual to lack
pigmentation, a condition called albinism.
The principal catabolic route for thyrosine involves first is transamination by thyrosine
aminotransferase, an enzyme in the liver whose level is regulated hormonally (Figure 21.21).
The product, p-hydroxyphenylpyruvate, is acted on by p-hydroxyphenylpruvatedioxygenase, an
unusual iron-containing enzyme, which catalyzes a ring hydroxylation, decarboxylation, and side
chain migration, using ascorbate as a cofactor, in a similar fashion to its use with
procollagenproline hydroxylase. This reaction involves a mechanism called the NIH shift, after
scientists at the national institute of health, who described a ring hydroxylation that proceeds via
formation of an epoxide intermediate.The phenylalanine hydroxylase reaction also involves an
NIH shift, with migration of the hydrogen at C-4 of phenylalanine to C-3 of thyrosine, without
mixing with solvent.

Figure 21.21. Catabolism of tyrosine to fumarate and acetoacetate

The product of p-hydroxyphenylpruvate oxidation, homogentisic acid, is oxidized by and
iron-containing enzyme, homogentisic acid dioxgenase, that cleaves the ring to yield a straightchain-eight-carbon compound that isomerizes to fumarylacetoacetate. The latter ultimately
cleves to yield fumarate and aceto-acetate, both of which are catabolized by standard energyyielding pathways. In plants, homogentisate is the precursor to the aromatic ring portion of
vitamin E.
A hereditary deficiency of the enzyme homogentisic acid dioxygenase in human causes a
condition that was known for centuries as the dark urine disease but is now called
alkaptunoria. Homogentisic acid accumulates and is excreted in large amounts in the urine; its

oxidation on standing causes the urine to become dark. Although the clinical symptoms of the
dieses are not severe,it is of considerable historical interest. Early in the twentieth century
Archibald garrod examined oedigrees of the families of afflicted individuals, and in 1908 he
proposed correctly that the deficiency was due to a gene controlling a normal reaction in the
breakdown of aromatic compounds. In other words, he proposed thet one gene encodes one
enzyme, long before the chemical nature of either genes or enzymes was known.
Tryptophan
Tryptophan is transformed by many pathways, of which just two concern us here-the
major catabolic route,proceeding via kynurenine to glutaryl-CoA ( Figure 21.22 ), and the
synthesis of nicotinamide nucleotides by a branch from this major catabolic pathway. The first
reaction in the degradation of tryptophan is catalyzed by tryptophan oxygenaase, an iron heme
protein whose level is controlled by two mechanisms: (1) induction by certain hormones and (2)
stabilization in vivo by its substrate, tryptophan, which increases enzyme levels by protecting the
protein against intracellular degradation.
NAD+ can be synthesized either from tryptophan or from the vitamin nicotinic acid (see
table 11.5 on page 389). For both pathways the last reaction is catalyzed by a glutaminerequiring amidotransferase( see margin). Both pathways probably contribute significantly to
NAD+ biosynthesis, as can be inferred from studies of the nicotinamide deficiency disease
pellagra. Pellagra was formerly endemic in regions, such as the sourthernunited states, where
corn is a dietary staple. Because cornmon proteins contain little tryptophan, deficiencies of this
amino acid where common. They symptoms of tryptophan deficiency are identical to those of
nicotinamidedeficienc, as expected if a major role of tryptophan is as a nicitinamide substitute.

Figure 21.22. Metabolic fates of tryptophan

Histidine
Histidene undergoes decarboxylation to generate histamine, a substance with murtiple
biological actions. When secreted in the stomach, histamine promotes the secretion of
hydrochloric acid and pepsin, both of which aid digestion. It is a potent vasodilator, released
locally in sites of trauma, inflammation, or allergic reaction. The local enlargement of blood
capillaries is the basis for the reddening that occurs in inflamed tissues. Release of histamine in
trauma contributes to the dangerous lowering of blood pressure that can lead to shock. A large
number of antihistamines are use to treat allergies and other inflammation. Typically, these drugs
prevent the binding of histamine to its receptors.

Figure 21.23 catabolism of histidine

Figure 21.23 presents the major catabolic route for histidine. This amino acid, like
trypthopan, does not undergo transamination at the start of its breakdown.rather, a specific lyase
cleaves out ammonia to give uronoic acid. Two subsequent steps bring about reduction and ring
opening to yield formiminoglutamicacid, which is of interest because it serves as a donor of
active one-carbon fragments. The formimino group is transferred to tetrahydrofolate, yielding 5formiminotertrahydrofolate and glutamate.
SERINE, GLYCINE, AND THREONINE
Although serine, glycine and threonine do not form a natural grouping, we consider them
together partly because of the hydroxyl group shared by serine, glycine and threonine and partly
because of the close interconnection between serine and glycine via the series
transhydrocymethylase reaction. Serine is quite active metabolically; we have already considered
its roles in biosynthesis is phospholipids and cysteine, as well as its contribution of activated
one-carbon units to the pool of tetrahydrofolate coenzyme. Glycine also plays murtiple roles,
including contributions to the one-carbon pool and as a precursor to glutathione, to purine
nucleotides, and to porphyrins. Figure 21.24 summarizes the metabolic fates of glycine and
serine. By contrast, threonine plays but one significant role other than as a constituent of
proteins. It is a precursor to isoleucine in plants and microorganism.

All three of these amino acids are synthesized and degraded by rather simple pathways.
Serine can be synthesized from glycine, via the serine transhydroxymethylase reaction. However,
this reaction probably proceeds more often in the reverse direction, a the principal biosynthetic
route to glycine and to activated one-carbon compounds. Most serine biosynthesis occurs in a
three-step sequence from the glycolytic intermediate 3-phosphoglycerate.

Threonine is an essential amino acid, with its synthesis limited to plants and prokaryotes.
Threonine synthesis begins with homoserine, which is derived froom aspartate. Homoserine
undergoes a phosphorylation, followed by a pyridoxal phosphate-dependent reaction that
eliminates phosphate and rehydrates the resultant double bond with hydroxyl group migration to
the -carbon.

The major route for glycine degradation is via the mitochondrial glycine cleavage system,
to yield NH3, CO2 and 5,10-methyleneterahydrofolate. This reaction particulary important in
photorespiration in plants. Glycolate produced in photorespiration is oxidized to glyoxylate,

which undergoes transmination to glycine. One molecule of glycine undergoes the glycine
cleavage reaction, and the resultant 5,10-methylenetetrahydrofolate reacts with a second
molecule of glycine, to give serine. Serine in turn is converted to 3-phosphoglycerate, which
undergoes the normal photosynthetic carbon reduction cycle.
Serine is catabolized both by its conversion to glycine and by the action of serinethreonine dehydratase. This pyridoxal phosphate-dependent enzyme converts serine to pyruvate
and threonine to -ketobutyrate (figure 21.25). -ketobutyrate is converted to propionyl-CoA and
CO2; the propionyl CoA is then metabolized to succinyl-CoA by the same pathways used in
odd-chain fatty acid oxidation and methionine catabolism. In plants and prokaryotes,

ketobutyrate has a biosynthetic role as well, as the first intermediate en route to isoleucine
biosynthesis.

Figure 21.25 reaction catalyzed by serine-threonine dehydratase

An alternative route for threonine catabolism is its reversible cleavage to glycine and
acetaldehyde, catalyzed by threonine aldolase. Though this reaction could in principle lead to
threonine biosynthesis in animals, acetaldehyde is highly toxic and cannot accumulate to the
point that it would be a biosynthesic intermediate.

VALINE, LEUCINE, ISOLEUCINE AND LYSINE

Valine, Leucine, Isoleucine and lysine have in common the fact that they are essential
amino acids for mammals, and they are synthesized primarily in plant and bacterial cells.
Furthermore, none of these amino acids is known to play significant metabolic roles other than as
protein constituents and as substrates for their own degradation. The pathways involved are
complex, and they are shown here only in outline.
Valine, Leucine and Isoleucine
Valine, Leucine and Isoleucine are structurally related, and they share certain reactions,
and enzymes in their biosynthetic pathways (figure 21.26). The last dour reaction in valine
biosynthesis and in isoleucine biosynthetesis are catalyzed by the same four enzyme. Valine
biosynthesis begins with transfer of a two-carbon fragment from hydroxyethyl thiamine
pyrophosphate to pyruvate. Similar transfer of a two-carbon unit to -ketobutyrate begins the
pathway to isoleucine. The keto acid analog of valine is the input for a four-step pathway to
leucine. In bacteria ecah of these three amino acids controls its own synthesis by feedback
inhibition of a different enzyme. In fact, the concept of allostreric control was developed largely
in studies on the inhibition of threonine dehydratase by isoleucine.

Figure 21.26 biosynthesis of valine and isoleucine

Degradation of leucine, isoleucine and valine in animals start with transamination followed by
oxidative decarboxylation of the respective keto acid.

The latter reaction is carried out by a multienzyme complex, called the branched, chain
keto acid dehydrogenese complex, which is similar in structure and mechanism to the pyruvate
dehydrogenease and - ketoglutarate dehydrogenase complexes. In a rare human disorder called
maple syrup urine disease, this complex is defective. All three keto acids accumulate in the urine,
and their characteristic odor gives the condition its name. The condition involves severe mental
retardation.
The remainder of the degradative pathways are comparable to fatty acid oxidation, except
that the intermediates must undergo a debranching not seen with -oxidation.
Lysine
Lysine metabolism is distinguished partly by the complexity of both its synthetic and degradative
pathways and partly by the fact that there are two distinct biosynthetic pathways. The
diaminophimelic acid pathway operate in bacteria, some lower fungi, algae and higher plants.

This pathways begins with condensation of pyruvate with aspartate

-semialdehyde and ends

with decarboxylation of diaminopimelate. The principle intermediate, diaminopimelate, also

serves an important fungtion as a constituent of bacterial cell walls. The

-aminoadipic acid

pathway is less widespread, functioning in other lower fungi, and in the protist Euglena.
PORPHYRIN AND HEME METABOLISM
BIOSYNTHESIS OF TETRAPYRROLES: THE SUCCINATE-GLYCINE PATHWAY
A major metabolic fate of glycine is its utilization for tetrapyrrole biosynthesis.
Tetrapyrole is generic term for compound containing four linkage pyrrole rings. Four such
classes of compounds are widespread in biology: the widely distributed iron porphyrin, heme;
the chlorophylls of plants and photosynthetic bacteria; the phycobilins, photosynthetic pigments
of algae (Chapter 17); and the cobalamins, notably vitamin B12 and its derivatives. Structures of
most of these compounds have been shown previously. All tetrapyrroles are synthesized from a
common precursor, -aminolevulinic acid (ALA). Figure 21.27 illustrates the relationship among
the various synhetic pathways.

We shall concentrate here on the well-understood porphyrin synthetic pathway, which lad
to heme. This pathway is widespread in animal tissue and, so far is known, is similar in all

organism containing heme proteins, such as cytochromes. Seven reactions are involved, and they
occur in two different cell compartments. The first reaction occurs in mitochondria, followed by
three reactions in the cytosol, and finally, three more mitochondrial reaction. As we shall see, this
compartmentation provides the opportunity for a novel control mechanism for the pathway.
Early labeling studies in animal revealed that all of the nitrogen of heme is deived from glycine,
and all of the carbon is derived from succinate and glycine. Hence, this synthesis is often called
the succinate-glycine pathway. The first reaction is catalyzed by a pyridoxal phosphatedependent enzyme, -aminolevulinic acid synthetase, or ALA synthetase. As shown in figure
21.28, the binding of glycine to pyridoxal phosphate probably activates the -carbon of glycine
for an attack on the thioester carbon of succinyl-CoA. Decarboxylation follow, to give product
ALA.

In plants, ALA is formed by a completely different pathway, a three step sequence

beginning with glutamate (Figure 21.29).

The first reaction of this novel pathway links glutamate, through its carboxyl group, to specific
transfer RNA, just as occurs in protein synthesis. The carboxyl group thus activated is then
reduced by NADPH, giving glutamate l-semialdehyde, which finally undergoes an internal
transamination to give ALA. Because the major end product of the pathway in plants is
chlorophyll, the synthesis ALA is regulated by light. The identity of the specific light-regulated
step is now under active investigation.
Whether in plants, animals, or microorganism, the reminder of the porphyrin synthetic
pathway involves three distinct process: (1) synthesis of a substituted pyrrole compound,
porphobilinogen from ALA; (2) condensation of four porphobilinogen molecules to yield a partly
reduced precursor called a phyrinogen; and (3) modification of the side chains, dehydrogenation,
of the ring system and introduction of iron, to give the porphyrin product, heme. In the first
stage, two molecules of ALA condense in the cytosol to form one molecule of porphobilinogen.
The reaction is catalyzed by ALA dehydratase:

Next, four molecules of porphobilinogen combine in a PLP-requiring deaminase reaction to give

the first tetrapyrrole compound, unroporphyrinogen III (Figure 21.30)

Two different proteins are involved in this reaction: uroporphyrinogen I synthase and
uroporphyrinogen III cosynthase. The former protein is enzymatically active by itself, but when
it acts alone, the product is the undesired symmetrical compound unroporphyrinogen I.
Interaction between the synthase and cosynthase allows one the rings to flip during the
combining reaction, so that the product is the asymmetric uroporphyrinogen III. The symmetric
compound and some metabolities derived from it asre synthesized as nonfunctionak side
products, in low amounts.
Uroporphyrinogen III undergoes decarboxylation of its acetic acid side chains. The
product then reenters the mitochondrion for further modification: first, side chain modification,
then ring oxidation to yield a fully conjugated system, and finally the insertion of iron. The last
reaction can proceed spontaneously, but it is catalyzed by ferrochelatase, an enzyme on the inner
mitochondrial membrane that also requires a reducing agent. At this stage the completed heme
combines with polypeptides to give completed heme protein, including myoglobin and
hemoglobin in vertebrates, and cytochromes and other heme proteins in all aerobic organism.
In a hereditary condition called congenital erythropoietic porphyria, uroporphyrinogen III
cosynthase is defective, and the symmetrical (and metabolically usesless) type I porphyrins
accumulate beyond the capacity of the body to excrete them. Their accumulation causes the urine
to turn red, the skin to become acutely photosensitive, and the teeth to become fluorescent, all
because of the deposition of the strongly light absorbing porphyrins. In addition, erythrocytes are
destroyed prematurely and insufficient heme is synthesized, maing afflicted individuals quite
anemic. It has been speculated that people labeled as vampires in medieval folktales suffered

from this condition, which would explain their performance for the dark, their bizarre
appearance, and their propensity for drinking blood. In fact, individuals with congenital
erythropoietic porphyria can be treated by injections of heme.
Quite distinct from the above condition is acute intermittent porphyria, which results
from deficiency in uroprophyrinogen I synthase. This deficiency causes ALA and
porphobilinogen to accumulate in the liver. The condition is accompanied by episodes of acute
abdominal pain and neurological disorders. There is evidence that King George III of England
suffered from this condition; evidence for this was put before the general public in the successful
stage play and film The Madness of King George. Symptoms of porphyrias can also be acquired,
most notably in lead poisoning. The crystal structure of ALA dehydratase show that lead can
displace zinc, the natural metal cofactor, and inhibit the enzyme, causing great accumulation of
ALA.
Being the first committed step in heme synthesis, the ALA synthetase reaction (Figure
21.28) is the major control point. Heme and related compounds feed-back-inhibit the enzyme.
Heme also has two other important effects. At low concentration, heme inhibit the synthesis of
ALA synthase at the translational level. At higher level, heme somehow blocks the translocation
of ALA synthetase from the cytosol, where it is synthesized on ribosomes into the
mitochondrion, where it acts. Heme also inhibits the ferrochelatase reaction (Figure 21.30). A
number of drugs and poisons cause excessive heme synthesis. In some cases,this effect result
from stimulation of the synthesis of cytochrome P450, whicj increase the demand for heme and
hence activates ALA synthetase.
Tetrapyrrole biosynthesis is being exploited as a target for the action of weed-killers. The
idea is to spray weeds in the dark with ALA. The pathway to chlorophyll begins, and when it
become light, the pathway is completed, and chlorophyll is produced in such massive amounts
that the plant weakness and dies.
DEGRADATION OF HEME IN ANIMAL
By far the most abundant porphyrin compound in vertebrates is the heme of hemoglobin.
Therefore, the story of porphyrin in degradation is largely the story of hemoglobin and heme
degradation. Lacking nuclei, mammalian erythroses are incapable of renewal and self-destruct
after characteristic interval. In humans the average erythroses life span is 120 days. Aged
erhtrocytes are destroyed upon passage through the spleen on liver (Figure 21.31).

Amino acids released from the globin portion of the hemoglobin molecule are catabolized
or reused for protein synthesis. The heme portion undergoes degradation, starting with a mixedfunction oxidase reaction that open the ring and converts one of the methane bridge carbon to
carbon monoxide. Iron is released from the resulting linear tetrapyrole, called biliverdin, and is
transported to storage pools in bone marrow for reuse in erythrocyte production. The tetrapyrrole
is next reduce to bilirubin, which is extracted. Bilirubin is quite insoluble, and its removal
involves several organ system. First, it complexes with serum albumin to transport to the liver.
There, bilirubin is solubilized by conjugation with two molecule of glucoronic acid. The reaction
is comparable to other glycol-syltransferase reaction we have encountered (see chapter 16) with
the substrate being UDP-glucuronate. This solubilized compound, bilirubin diglucuronide, is
secreted into the bile and ultimately excreted via the intestine.
Because several organ system participate in the degradation of heme, there are numerous
ways for things to go wrong. When heme catabolism is defective, bilirubin accumulates in the
blood. This defect is first recognize because the distinctive color of bilirubin gives yellow cast to
the skin and the white of the eyes. This condition, known as jaundice, is seen, for example, in
acute or chronic liver disease, in which the glucuronate conjugating system is impaired and
albumin as shown in figure 21.32, the pathway to catecholamines is similar, starting with a
tetrahydrobopetrin, dependent hydroxylation (of tyrosine) followed by decarboxylation. The
hydroxylation product is dopa, which formed by a quite different mechanism in melanine
synthase. However, the latter pathways is localized to the melanocytes, whereas most
catecholamine synthesis occurs in the adrenal medulla and in the central nervous system.

Once formed, dopa undergoes decarbocylation to give dopamine. Dopamine serves in

turn as substrate for a copper-containing monooxygenase, dopamine -hydroxylase, giving
norepinephrine, which in turn is methylated by S-adenosyl-methionine to give epinephrine.
Although dopamine and norepinephrine are intermediates in epinephrine synthesis, each is a
neurotransmitter in its own right.
BIOCHEMISTRY OF NEUROTRANSMISSION
In chapter 10 we noted that the transmission of nerve impulses involve two distinc components:
(1) the transmission of an action potential within a neuron by continuous polarization and
depolarization of the membrane; and (2) transmission of the impulse across synaptic junction,

from one neuron to another neuron or to a muscle or gland cell. Transmission within a neuron is
described in chapter 10. Here we discuss transmission from cell to cell.
The Cholinergic Synapse
Neurontransmission usually involves the release of a chemical messenger, the
neurontransmitter, from the presynaptic cell and its binding to receptors on the post-synaptic cell
(nerve, muscle, or gland). Synapses involving acetylcohalines as the neurotransmitter
(cholinergic synapses) are the best understood. Recall from Chapter 19 that choline is
synthesized principally as part of phosphatidylcoline, with the carbon of choline ultimately of
serine.
The Nicotinic Acethylcholine Receptor
Viewed directly from the surface, the nicotinic acethylcholoine receptor appear in the
electron microscope to be doughnutlike structure extending through the membrane and
containing a central pore (figure 21.34).

Isolation of the receptor by affinity chromatography showed them to be pentamers made of four
kinds of glycoprotein subunit (molecular masses 54, 56, 58 and 60 kilodaltons in molar ratio
2:1:1:1). Reconstitution of receptor into lipid visicles providesa system that can conduct ions
when stimulated by acethylcoline. Thus, it appears that the receptor and ion chanel are single
unit. The central pore presumably functions as the gated ion channel. Genes for many of the
components of the synaptic system have been cloned, including choline acethyltrasferase,
acethylcholinesterase and components of the nicotinic reseptors.
Although postsynaptic membranes are typically packed densely with receptors (of the
order of 20,000 per square micrometer), these region constitute only a small fraction of the
cellular surface in a typical neural tissue. Therefore, biochemist often turn to more specialized
tissues for study of synaptic processes and biochemistry. Favorite experimental subjects for such

studies are the electric organ of the electric ray (Torpedo) and the electric eel (Electrophorus).
These organ contain stack of cells called electropaques, which have a high density (105/m2) of
nicotinic achetylcholin receptors extending over one whole of the face of the cell. Depolarization
of the membrane on this face, while the other face remain at the resting potential, give a of
130 mV across the cell. With thousands of cells stacked in series, potentials of several hundred
volts are generated.
Inhibition of the Cholinergic Synaptic Transmission
Achetylcholinesterase is a serine esterase and consequently can be irreversibly inhibited
by reagents that react with the active site of serine, such us diisopropyl fluorophosphates, sarin,
physostigmine, and parathion. As you might expect, acethylcholinesterase inhibitors are
extremely toxic substances, causing paralysis. Another class of toxins acts on the acethylcholine
receptors itself either by blocking it (d-tubocurarine, from curare, and the small protein toxins in
some snake venoms, like cobra toxin) or by blocking the ion channels open (nicotine). The
former class are called antagonists, and the latter agonists. Such compound are useful, both as
research tools and, with proper caution, as drugs (muscle relaxants).
Catecholamines and Adrenergic Neurons
We concentrated on the cholinergic synapse because it is the best understood at present.
But many other substances are known suspected to be neutranssmitters, which function in
different kinds of synapse. These include the catecholiamines-dopamine, norepinephrine and
epinephrine. Because epinephrine is also an adrenal hormone, synapse that use catecholamines
are called adrenergic.
The importance of dopamine in neutral transmission is emphasized by the number of
major neurological diseases that are associated with improper dopamine regulation. The earliest
indication of this type of defect was the finding that dopamine levels are abnormally low in
particular region of the brain of patients with parkinsonism, a severe neurological disorder.
Attempts to threat such patients with dopamine were futile, because this substance, after
injection, does not cross the blood-brain barrier. For many individuals with parkinsonism, daily
doses of dopa have provided dramatic clinical improvement.
Circumstantial evidence links dopamine metabolism to schizophrenia, which may result
partly from excess firing of dopaminergic neurons-neurons secreting dopamine. Part of

circumstantial evidence is the close structural relationship between dopamine and mescaline, a
product of the peyote cactus known to induce a quasi schizophrenic state. Amphetamine is
another

catecholamine

analog

with

potent

physocopharmological

properties.

Other

circumstantial evidence is that the drugs most useful for treating schizophrenia, notably
chlorpromazine, are dopamine antagonists, which block receptors have been cloned, and intens
interest is focus upon which, if any, of these receptors fuctions abnormally in schizophrenia.

Another psychotropic drug, whose action suggested a role for serotonin in

neurotransmission, is lysergic acid diethylamide (LSD), an indole derivative. LSD action is
explained in a part as mimicking the effect of serotonin at receptors in the central nervous
system. More recently the drug fluoxetine (marked as Prozac) has been showed to block the
uptake of serotonin into presynaptic nerve terminals, with an increase in the amount available to
bind to postsynaptic receptors. Originally marketed as an antidepressant drug, Prozac is now
known to be active against a range psychiatric disorders.

Also related to serotonin metabolism is another widely prescribed drug, Ritalin, used to
treat children with hyperactivity-attention deficit disorder. Because Ritalin is stimulant, now to
act by raising dopamine levels, its effect in calming hyperactive children was paradoxical.
Studies reported in early 1999, however, reported that the drug, in the low doses used with such
children, actually exerts its calming effect by elevating levels of serotonin.
Just as acethylcholinesterase limit the firing of cholinergic synapse, so also do two
enzymes regulate the action of adrenergic neurons, these enzyme are catecholamine Omethyltransferase (COMT), which catalyze an AdoMet-dependent transmethylation, and
monoamine oxidase (MAO), a flavoprotein that oxidizes primary amines to aldehydes.
Excitatory and Inhibitory Neurotransmission
The necessity for many types of neurotransmitters and corresponding kinds of synapse
probably derives from the complexity of vertebrate neural system. Neurotransmitters and the
synapses that they mediate vary widely in their properties. Some are rapid in their action, and
some are slow. Some, such as the nicotinic cholinergic synapses and thus involving glutamate,
are stimulatory and promote an action potential in the postsynaptic cell. Others, such as those
using -aminobutyric acid (GABA) as a transmitter, are inhibitory an impulse revieved as these
synapses will discourage the transmitter of an action potential in the recipient neuron. This
inhibition can occur, for example, by opening chloride channel. Weather the neuron fire depend
on the net summation of stimulatory and inhibitory input. Given that GABA, a principle
inhibitory neurotransmitter, is derived from glutamate, it is interesting that glutamate itself is a
principle excitatory neutransmitter. Glutame ingested in large quantities, as the flavor enhancer
monosodium glutamate (MSG), can damage the central nervous system by overstimulation. For

this reason, MSG was removed from most infant formulas two decades ago, because the
developing nervous system is particularly susceptible to damage. MSG is a major constituent of
soy souce, and some people experience sweetnest, flushing and headaches from eating MSGflavored foods.
A second type of receptor for achetylcholine, the muscarinic acethylcholine receptor, is
quite different. This receptor which has different structure can be inhibitory, though not by the

chloride channel mechanism. Thus, weather acetylcholine is stimulatory or inhibitory depends on

the type of receptor to which it binds. Inhibitory synapses play a major role in the regulation of
neural transmission. A typical nerve input from the axons of many different neurons. Figure
below is schematic illustration of transmission and inhibition in a neural network.

Multiple synapses on the body of a single neuron

Certain small peptides, such as somatostatin, neurotensin, and the enkephalins, also act as
neurotransmitter (shown on the table below). In some cases this compounds exihibit a second
function as neurohormones, which are describe in the last section.
Finally, there are even specialized synapses that do not use neurotransmitters substances
at all. Although transmission through a synapse via neurotransmitter can be quite rapid (about a
millisecond), some responses must be even quicker than this will allow. In such cases, there is
direct electrical ionic conduction between neural cells, using gap junction. Such electrical
synapses are frequently found in animal that live in cold environments yet need to make quick
motion. The low body temperatures of such creatures would slow the chemical and diffusion
processes involved in chemical synapses. Direct conduction provides a solution to this problem.
Neurotransmitter Receptors and Psychopharmacology
Recent years have seen explosive growth in our understanding of relationship between
particular neurotransmitters and receptors, on the one hand, and specific behavior patterns and
abnormalities, on the other including aggressive behavior, anxiety, biochemical changes
accompanying learning, and phsychiatric disorders such as schizophrenia and drug addiction.
Most of our enlarged understanding has emerged from the discovery of multiple receptor classes,
such as the four distinct dopamine receptors mention earlier. This has permitted two lines of
attack: (1) psychopramalogical approach, in which studies on recombinant receptor properties
are allowing the development of agonist and antagonist specific to one or two receptror subtypes,

for studies in both experimental animal and human and (2) molecular genetic approaches, in
which the development of transgenic animal models allows the selective deletion of the gene for
a particular subtype in an experimental animal, usually a :knockout mouse.
Table of Some peptides that act as neurohormones (H) or neurotransmitter (T)
Name
H/T
Sequence
-Endorphin
H
YGGFMTSFKSQTPLVTLFKNAYKKGE
Met-enkephalin
H,T
YGGFM
Leu encephalin
H,T
YGGFL
Neurotensin
T
pELYENKPRRPYIL
Somatostatin
T
AGCKNFFWKTFTSC
Current research on the biochemistry of drug addiction provides good examples of what
is being learned about molecular control of behavior. It is a long been suspected that dopamine
metabolism is intimately connected with addiction, because all addictive drug have been shown
to cause a surge in dopamine levels in the reward section of brain, the nucleus accumbens. In one
study, agonists for the closely related D1 and D2 dopamine receptors in rats were found to exert
opposite effects upon the animals tendency to dose themselves with cocaine after an initial drug
experience. In another study a targeted deletion of the D4 receptor in mise cause them to become
hypersensitive to ethanol, cocaine, and methamphetamine.
Recent work, however, has implicated glutamate receptors as well as in control addictive
behavior. Inhibition of glutamate neurotransmission in rats has been shown to modulate the
compulsive drug seeking behavior after an initial drug experience, and neuroscientist are
excited at the prospect of developing therapies involving glutamate antagonist, which might
increase the likelihood that an addict would remain clean after treatment. Glutamate receptor also
strongly linked to the action of phencyclidine (PCP, or angle dust). This compound blocks
glutamate binding to the N-methyl_D_aspartate (NMDA) class of glutamate receptor, thereby
inducing a schizophrenia like state though until recently to result from decreased glutamergic
neutransmission. However, neuroscientist have found that lowering brain glutamate levels with
another drug greatly dimishes the effectiveness of PCP, suggesting a possible new approach to
the treatment of schizophrenia.
Glutamate receptor are receving greatly increase attention in other regards. A particular
receptor subtype has been shown to be phosphorylated by a calmodulin dependent kinase during

long term potentiation, a neurophysiological response associated with learning, suggesting

that this covalent receptor modification might be associated with long term memory.
Neurohormones
Whereas transmitter substances carry out the actual passage of a signal across a synapse, other
substances in nervous systems modify the way in which nerve cells respond to transmitters.
These substances include peptides called neurohormones, which were discovered originally as a
result of Solomon Synders studies on addiction to opiate drugs such as morphine. Synder
detected receptor in brain tissue specific for binding opiates. Because it seemed unlikely that
vertebrate brains would contain specific receptors directed toward a product of the poppy plant,
as search was made for the natural ligands that bound to these sites. The search led, in the 1970s,
to the finding of several small peptides, called enkephalins and endorphins, which are natural
analgesics. The modification of neural signal by these substances appears to be responsible for
the insensitivity to pain that is experiences under condition of great stress or shock. The
effectiveness of opiate analgesics such as morphine is as consequence, perhaps accidental, of the
recognition of these opiates by neurohormone receptor despite their structural differences from
neurohormones.
The endorphins and enkephalins are synthesized as a part of the much longer hormone
precursor prepro opiomelanocortin. As describe in the other chapter, this precursor is cleaved
to release both the neurohormones and a number of other hormones with entirely different
function.