S E C T I O N

Metabolism of Proteins &

VI Amino Acids

Biosynthesis of the C H A P T E R

Nutritionally Nonessential
Amino Acids
Victor W. Rodwell, PhD
OBJ E C TI VE S
After studying this chapter,
you should be able to:
27
■ Explain why the absence of certain amino acids that are present in most
proteins from the human diet typically is not deleterious to human health.
■ Appreciate the distinction between the terms “essential” and “nutritionally
essential” amino acids, and identify the amino acids that are nutritionally
nonessential.
■ Name the intermediates of the citric acid cycle and of glycolysis that are
precursors of aspartate, asparagine, glutamate, glutamine, glycine, and serine.
■ Illustrate the key role of transaminases in amino acid metabolism.
■ Explain the process by which the 4-hydroxyproline and 5-hydroxylysine of
proteins such as collagen are formed.
■ Describe the clinical presentation of scurvy, and provide a biochemical
explanation for why a severe deprivation of vitamin C (ascorbic acid) results in
this nutritional disorder.
■ Appreciate that, despite the toxicity of selenium, selenocysteine is an essential
component of several mammalian proteins.
■ Define and outline the reaction catalyzed by a mixed-function oxidase.
■ Identify the role of tetrahydrobiopterin in tyrosine biosynthesis.
■ Indicate the role of a modified transfer RNA (tRNA) in the cotranslational
insertion of selenocysteine into proteins.

BIOMEDICAL IMPORTANCE significant nutritiona and metaboic abnormaities. Both the

Amino acid deficiency states can resut if nutritionay essentia
amino acids are absent from the diet, or are present in inade-
quate amounts. In certain regions of West Africa, for exampe,
kwashiorkor sometimes resuts when a chid is weaned onto
a starchy diet poor in protein, whie marasmus may occur
if both caoric intake and specific amino acids are deficient.
Patients with short bowe syndrome, if unabe to absorb suf-
ficient quantities of caories and nutrients, may suffer from
nutritiona disorder scurvy, a dietary deficiency of vitamin C,
and specific genetic disorders are associated with an impaired
abiity of connective tissue to form peptidy 4-hydroxyproine
and peptidy 5-hydroxyysine. The resuting conformationa
instabiity of coagen is accompanied by beeding gums,
sweing joints, poor wound heaing, and utimatey in death.
Menkes syndrome, characterized by kinky hair and growth
retardation, resuts from a dietary deficiency of copper, an
essentia cofactor for the enzyme ysy oxidase that functions

273
274 SECTION VI Metabolism of Proteins & Amino Acids

in formation of the covaent cross-inks that strengthen coa-
gen fibers. Genetic disorders of coagen biosynthesis incude
severa forms of osteogenesis imperfecta, characterized by
fragie bones, and Ehlers-Danlos syndrome, a group of con-
nective tissue disorders that resut in mobie joints and skin
abnormaities due to defects in the genes that encode enzymes,
incuding procoagen-ysine 5-hydroxyase.
NUTRITIONALLY ESSENTIAL &
NUTRITIONALLY NONESSENTIAL
AMINO ACIDS
Whie often empoyed with reference to amino acids, the terms
“essentia” and “nonessentia” are miseading, since for human
subjects a 20 common amino acids are essentia to ensure
heath. Of these 20 amino acids, 8 must be present in the human
diet, and thus are best termed “nutritionally essential.” The other
12 “nutritionally nonessential” amino acids, whie metabolically
essentia, need not be present in the diet (Table 27–1). The dis-
tinction between these two casses of amino acids was estabished
in the 1930s by feeding human subjects a diet in which purified
amino acids repaced protein. Subsequent biochemica investiga-
tions utimatey reveaed the reactions and intermediates invoved
in the biosynthesis of a 20 amino acids. Amino acid deficiency
disorders are endemic in certain regions of West Africa where
diets rey heaviy on grains that are poor sources of tryptophan
and ysine. These nutritiona disorders incude kwashiorkor,
which resuts when a chid is weaned onto a starchy diet poor in
protein, and marasmus, in which both caoric intake and spe-
cific amino acids are deficient.
Biosynthesis of the Nutritionally
Essential Amino Acids Involves
Lengthy Metabolic Pathways
The existence of nutritiona requirements suggests that depen-
dence on an external source of a specific nutrient can be of
greater surviva vaue than the abiity to biosynthesize it. Why?
Because if the diet contains ampe quantities of a nutrient,
retention of the abiity to biosynthesize it represents informa-
tion of negative surviva vaue, because ATP and nutrients are
not required to synthesize “unnecessary” DNA—even if spe-
cific encoded genes are no onger expressed. The number of
enzymes required by prokaryotic ces to biosynthesize the
nutritionay essential amino acids is arge reative to the num-
ber of enzymes required for the formation of the nutritionay
nonessential amino acids (Table 27–2). This suggests a sur-
viva advantage for humans to retain the abiity to biosynthe-
tize “easy” amino acids whie osing the abiity to make others
more difficut to biosynthesize. The reactions by which organ-
isms such as pants and bacteria, but not human subjects, form
certain amino acids are not discussed. This chapter addresses
the reactions and intermediates invoved in the biosynthesis of
the 12 nutritionay nonessential amino acids in human tissues
and emphasizes seected medicay significant disorders asso-
ciated with their metaboism.
TABLE 27–2 Enzymes Required for the Synthesis of
Amino Acids From Amphibolic Intermediates
Number of Enzymes Required to Synthesize

Nutritionally Essential Nutritionally Nonessential

a
Arg 7 Ala 1
TABLE 27–1 Amino Acid Requirements of Humans
His 6 Asp 1
Nutritionally Essential Nutritionally Nonessential
b
Thr 6 Asn 1
Argininea Alanine
Met 5 (4 shared) Glu 1
Histidine Asparagine
a
Lys 8 Gln 1
Isoleucine Aspartate
c
Ile 8 (6 shared) Hyl 1
Leucine Cysteine
d
Val 6 (all shared) Hyp 1
Lysine Glutamate
a
Leu 7 (5 shared) Pro 3
Methionine Glutamine
Phe 10 Ser 3
Phenylalanine Glycine
e
Trp 5 (8 shared) Gly 1
Threonine Hydroxyprolineb
f
59 (total) Cys 2
Tryptophan Hydroxylysineb
g
Tyr 1
Valine Proline
17 (total)
Serine
a
From Glu.
Tyrosine b
From Asp.
c
From Lys.
a d
Nutritionally “semiessential.” Synthesized at rates inadequate to support growth of From Pro.
e
children. From Ser.
b f
Not necessary for protein synthesis, but is formed during posttranslational process- From Ser plus sulfate.
g
ing of collagen. From Phe.
 CHAPTER 27 Biosynthesis of the Nutritionally Nonessential Amino Acids 275

NH3+
–
O3PO O–

O O

FIGURE 27–3 γ-Glutamyl phosphate.

Glutamate Dehydrogenase, Glutamine

FIGURE 27–1 The reaction catalyzed by glutamate dehydro-
genase (EC 1.4.1.3).
BIOSYNTHESIS OF THE
NUTRITIONALLY NONESSENTIAL
AMINO ACIDS
Glutamate
Gutamate, the precursor of the so-caed “gutamate famiy” of
amino acids, is formed by the reductive amidation of the citric
acid cyce intermediate α-ketogutarate, a reaction catayzed
by mitochondria gutamate dehydrogenase (Figure 27–1).
The reaction both strongy favors gutamate synthesis and
owers the concentration of cytotoxic ammonium ion.
Glutamine
The amidation of gutamate to gutamine catayzed by gutamine
synthetase (Figure 27–2) invoves the intermediate formation of
γ-gutamy phosphate (Figure 27–3). Foowing the ordered bind-
ing of gutamate and ATP, gutamate attacks the γ-phosphorus of
ATP, forming γ-gutamy phosphate and ADP. NH4+ then binds,
and uncharged NH3 attacks γ-gutamy phosphate. Reease of Pi
and of a proton from the γ-amino group of the tetrahedra inter-
mediate then aows reease of the product, gutamine.
Alanine & Aspartate
Transamination of pyruvate forms aanine (Figure 27–4). Simi-
ary, transamination of oxaoacetate forms aspartate.
Synthetase, & Aminotransferases
Play Central Roles in Amino Acid
Biosynthesis
The combined action of the enzymes gutamate dehydrogenase,
gutamine synthetase, and the aminotransferases (see Figures 27–1,
27–2, and 27–4) resuts in the incorporation of potentiay cyto-
toxic ammonium ion into nontoxic amino acids.
Asparagine
The conversion of aspartate to asparagine, catayzed by aspara-
gine synthetase (Figure 27–5), resembes the gutamine syn-
thetase reaction (see Figure 27–2), but gutamine, rather than
ammonium ion, provides the nitrogen. Bacteria asparagine
synthetases can, however, aso use ammonium ion. The reac-
tion invoves the intermediate formation of asparty phosphate
(Figure 27–6). The couped hydroysis of PPi to Pi by pyro-
phosphatase, EC 3.6.1.1, ensures that the reaction is strongy
favored.
Serine
Oxidation of the α-hydroxy group of the gycoytic interme-
diate 3-phosphogycerate, catayzed by 3-phosphogycerate
dehydrogenase, converts it to 3-phosphohydroxypyruvate.
Transamination and subsequent dephosphoryation then
form serine (Figure 27–7).
Glycine
Gycine aminotransferases can catayze the synthesis of gy-
cine from gyoxyate and gutamate or aanine. Unike most
aminotransferase reactions, is strongy favored gycine syn-
thesis. Additiona important mammaian routes for gycine
formation are from choine (Figure 27–8) and from serine
(Figure 27–9).

NH3+ NH3+
–
O O– H2N O–

O O O O
L-Glutamate L-Glutamine

NH4+

Mg-ATP Mg-ADP + Pi
FIGURE 27–4 Formation of alanine by transamination of
FIGURE 27–2 The reaction catalyzed by glutamine synthetase pyruvate. The amino donor may be glutamate or aspartate. The other
(EC 6.3.1.2). product thus is α-ketoglutarate or oxaloacetate.
276 SECTION VI Metabolism of Proteins & Amino Acids

+ +
O NH 3 O NH 3
– –
O O
–
O H 2N
O O
L-Aspartate L-Asparagine

H2O + Gln Glu

Mg-ATP Mg-AMP + PPi

FIGURE 27–5 The reaction catalyzed by asparagine synthe-

tase (EC 6.3.5.4). Note similarities to and differences from the gluta-
mine synthetase reaction (Figure 27–2).

FIGURE 27–7 Serine biosynthesis. Oxidation of

Proline 3-phosphoglycerate is catalyzed by 3-phosphoglycerate dehydroge-
The initia reaction of proine biosynthesis converts the nase (EC 1.1.1.95). Transamination converts phosphohydroxypyruvate
γ-carboxy group of gutamate to the mixed acid anhydride of to phosphoserine. Hydrolytic removal of the phosphoryl group cata-
lyzed by phosphoserine hydrolase (EC 3.1.3.3) then forms l-serine.
gutamate γ-phosphate (see Figure 27–3). Subsequent reduc-
tion forms gutamate γ-semiadehyde, which foowing spon-
taneous cycization is reduced to l-proine (Figure 27–10).

Cysteine
Whie not itsef nutritionay essentia, cysteine is formed from
methionine, which is nutritionay essentia. Foowing con-
version of methionine to homocysteine (see Figure 29–18),
homocysteine and serine form cystathionine, whose hydroy-
sis forms cysteine and homoserine (Figure 27–11).

Tyrosine
Phenyaanine hydroxyase converts phenyaanine to tyro-
sine (Figure 27–12). If the diet contains adequate quantities
of the nutritionay essentia amino acid phenyaanine, tyrosine
is nutritionay nonessentia. However, since the phenyaanine
hydroxyase reaction is irreversibe, dietary tyrosine cannot
repace phenyaanine. Cataysis by this mixed-function oxidase
incorporates one atom of O2 into the para position of phenyaa-
nine and reduces the other atom to water. Reducing power, pro-
vided as tetrahydrobiopterin, derives utimatey from NADPH.

Hydroxyproline & Hydroxylysine FIGURE 27–8 Formation of glycine from choline. Catalysts
Hydroxyproine and hydroxyysine occur principay in coagen. include choline dehydrogenase (EC 1.1.3.17), betaine aldehyde dehy-
Since there is no tRNA for either hydroxyated amino acid, nei- drogenase (EC 1.2.1.8), betaine-homocysteine N-methyltransferase
(EC 2.1.1.157), sarcosine dehydrogenase (EC 1.5.8.3), and dimethylgly-
ther dietary hydroxyproine nor dietary hydroxyysine is incor- cine dehydrogenase (EC 1.5.8.4).
porated during protein synthesis. Peptidy hydroxyproine and
hydroxyysine arise from proine and ysine, but ony after these Methylene
amino acids have been incorporated into peptides. Hydroxyation H4 folate H4 folate
of peptidy proy and peptidy ysy residues, catayzed by prolyl NH3+ NH3+
hydroxylase and lysyl hydroxylase of skin, skeeta musce, and O– O–

+
HO O O
O NH 3
– Serine H2O Glycine
O
–O PO
3
O FIGURE 27–9 Interconversion of serine and glycine, cata-
lyzed by serine hydroxymethyltransferase (EC 2.1.2.1). The reac-
FIGURE 27–6 Aspartyl phosphate. tion is freely reversible. (H4 folate, tetrahydrofolate.)
 CHAPTER 27
FIGURE 27–10 Biosynthesis of proline from glutamate.
Catalysts for these reactions are glutamate-5-kinase (EC 2.7.2.11),
glutamate-5-semialdehyde dehydrogenase (EC 1.2.1.41), and pyrroline-
5-carboxylate reductase (EC 1.5.1.2). Ring closure of glutamate semi-
aldehyde is spontaneous.
granuating wounds requires, in addition to the substrate, moec-
uar O2, ascorbate, Fe2+, and α-ketogutarate (Figure 27–13).
For every moe of proine or ysine hydroxyated, one moe of
α-ketogutarate is decarboxyated to succinate. The hydroxyases
are mixed-function oxidases. One atom of O2 is incorporated into
proine or ysine, the other into succinate (see Figure 27–13). A
deficiency of the vitamin C required for these two hydroxyases
resuts in scurvy, in which beeding gums, sweing joints, and
impaired wound heaing resut from the impaired stabiity of co-
agen (see Chapters 5 and 50).
Biosynthesis of the Nutritionally Nonessential Amino Acids 277
FIGURE 27–11 Conversion of homocysteine and serine to
homoserine and cysteine. The sulfur of cysteine derives from methio-
nine and the carbon skeleton from serine. The catalysts are cystathionine
β-synthase (EC 4.2.1.22) and cystathionine γ-lyase (EC 4.4.1.1).
Valine, Leucine, & Isoleucine
Whie eucine, vaine, and isoeucine are a nutritionay essentia
amino acids, tissue aminotransferases reversiby interconvert
a three amino acids and their corresponding α-keto acids.
FIGURE 27–12 Conversion of phenylalanine to tyrosine by
phenylalanine hydroxylase (EC 1.14.16.1). Two distinct enzymatic
activities are involved. Activity II catalyzes reduction of dihydrobiop-
terin by NADPH, and activity I the reduction of O2 to H2O and of phe-
nylalanine to tyrosine. This reaction is associated with several defects
of phenylalanine metabolism discussed in Chapter 29.
278 SECTION VI Metabolism of Proteins & Amino Acids
FIGURE 27–13 Hydroxylation of a proline-rich peptide.
Molecular oxygen is incorporated into both succinate and proline.
Procollagen-proline 4-hydroxylase (EC 1.14.11.2) thus is a mixed-
function oxidase. Procollagen-lysine 5-hydroxylase (EC 1.14.11.4)
catalyzes an analogous reaction.
These α-keto acids thus can repace their corresponding amino
acids in the diet.
Selenocysteine, the 21st Amino Acid
Whie the occurrence of seenocysteine (Figure 27–14) in pro-
teins is reativey uncommon, at east 25 human seenoproteins
are known. Seenocysteine is present at the active site of severa
human enzymes that catayze redox reactions. Exampes incude
thioredoxin reductase, gutathione peroxidase, and the deiodinase
that converts thyroxine to triiodothyronine. Where present,
seenocysteine participates in the cataytic mechanism of these
enzymes. Significanty, the repacement of seenocysteine by
cysteine can actuay reduce cataytic activity. Impairments in
human seenoproteins have been impicated in tumorgenesis
and atheroscerosis, and are associated with seenium deficiency
cardiomyopathy (Keshan disease).
Biosynthesis of seenocysteine requires serine, seenate
(SeO42−), ATP, a specific tRNA, and severa enzymes. Serine pro-
vides the carbon skeeton of seenocysteine. Seenophosphate,
formed from ATP and seenate (see Figure 27–14), serves as the
seenium donor. Unike 4-hydroxyproine or 5-hydroxyysine,
seenocysteine arises cotranslationally during its incorporation
into peptides. The UGA anticodon of the unusua tRNA caed
tRNASec normay signas STOP. The abiity of the protein syn-
thetic apparatus to identify a seenocysteine-specific UGA
codon invoves the seenocysteine insertion eement, a stem-
oop structure in the untransated region of the mRNA. tRNASec
is first charged with serine by the igase that charges tRNASer.
H
H Se CH2 C COO–
NH3+
O
Se + ATP + H2O AMP + Pi + H Se P O–
O –
FIGURE 27–14 Selenocysteine (top) and the reaction cata-
lyzed by selenophosphate synthetase (EC 2.7.9.3) (bottom).
Subsequent repacement of the serine oxygen by seenium
invoves seenophosphate formed by seenophosphate synthe-
tase (see Figure 27–14). Successive enzyme-catayzed reactions
convert cystey-tRNASec to aminoacryy-tRNASec and then to
seenocystey-tRNASec. In the presence of a specific eongation
factor that recognizes seenocystey-tRNASec, seenocysteine
can then be incorporated into proteins.
SUMMARY
■ A vertebrates can form certain amino acids from amphiboic
intermediates or from other dietary amino acids. The
intermediates and the amino acids to which they give rise are
α-ketogutarate (Gu, Gn, Pro, Hyp), oxaoacetate (Asp, Asn),
and 3-phosphogycerate (Ser, Gy).
■ Cysteine, tyrosine, and hydroxyysine are formed from
nutritionay essentia amino acids. Serine provides the carbon
skeeton and homocysteine the sufur for cysteine biosynthesis.
■ In scurvy, a nutritiona disease that resuts from a deficiency
of vitamin C, impaired hydroxyation of peptidy proine and
peptidy ysine resuts in a faiure to provide the substrates for
cross-inking of maturing coagens.
■ Phenyaanine hydroxyase converts phenyaanine to tyrosine.
Since the reaction catayzed by this mixed function oxidase is
irreversibe, tyrosine cannot give rise to phenyaanine.
■ Neither dietary hydroxyproine nor hydroxyysine is
incorporated into proteins because no codon or tRNA dictates
their insertion into peptides.
■ Peptidy hydroxyproine and hydroxyysine are formed
by hydroxyation of peptidy proine or ysine in reactions
catayzed by mixed-function oxidases that require vitamin C as
cofactor.
■ Seenocysteine, an essentia active site residue in severa
mammaian enzymes, arises by cotransationa insertion from a
previousy modified tRNA.
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