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
27
OBJ E C TI VE S ■ 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.
After studying this chapter, ■ Appreciate the distinction between the terms “essential” and “nutritionally
you should be able to: 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

nutritiona disorder scurvy, a dietary deficiency of vitamin C,
Amino acid deficiency states can resut if nutritionay essentia and specific genetic disorders are associated with an impaired
amino acids are absent from the diet, or are present in inade- abiity of connective tissue to form peptidy 4-hydroxyproine
quate amounts. In certain regions of West Africa, for exampe, and peptidy 5-hydroxyysine. The resuting conformationa
kwashiorkor sometimes resuts when a chid is weaned onto instabiity of coagen is accompanied by beeding gums,
a starchy diet poor in protein, whie marasmus may occur sweing joints, poor wound heaing, and utimatey in death.
if both caoric intake and specific amino acids are deficient. Menkes syndrome, characterized by kinky hair and growth
Patients with short bowe syndrome, if unabe to absorb suf- retardation, resuts from a dietary deficiency of copper, an
ficient quantities of caories and nutrients, may suffer from 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- Biosynthesis of the Nutritionally
gen fibers. Genetic disorders of coagen biosynthesis incude
severa forms of osteogenesis imperfecta, characterized by
Essential Amino Acids Involves
fragie bones, and Ehlers-Danlos syndrome, a group of con- Lengthy Metabolic Pathways
nective tissue disorders that resut in mobie joints and skin The existence of nutritiona requirements suggests that depen-
abnormaities due to defects in the genes that encode enzymes, dence on an external source of a specific nutrient can be of
incuding procoagen-ysine 5-hydroxyase. 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-
NUTRITIONALLY ESSENTIAL & tion of negative surviva vaue, because ATP and nutrients are
NUTRITIONALLY NONESSENTIAL not required to synthesize “unnecessary” DNA—even if spe-
AMINO ACIDS cific encoded genes are no onger expressed. The number of
enzymes required by prokaryotic ces to biosynthesize the
Whie often empoyed with reference to amino acids, the terms nutritionay essential amino acids is arge reative to the num-
“essentia” and “nonessentia” are miseading, since for human ber of enzymes required for the formation of the nutritionay
subjects a 20 common amino acids are essentia to ensure nonessential amino acids (Table 27–2). This suggests a sur-
heath. Of these 20 amino acids, 8 must be present in the human viva advantage for humans to retain the abiity to biosynthe-
diet, and thus are best termed “nutritionally essential.” The other tize “easy” amino acids whie osing the abiity to make others
12 “nutritionally nonessential” amino acids, whie metabolically more difficut to biosynthesize. The reactions by which organ-
essentia, need not be present in the diet (Table 27–1). The dis- isms such as pants and bacteria, but not human subjects, form
tinction between these two casses of amino acids was estabished certain amino acids are not discussed. This chapter addresses
in the 1930s by feeding human subjects a diet in which purified the reactions and intermediates invoved in the biosynthesis of
amino acids repaced protein. Subsequent biochemica investiga- the 12 nutritionay nonessential amino acids in human tissues
tions utimatey reveaed the reactions and intermediates invoved and emphasizes seected medicay significant disorders asso-
in the biosynthesis of a 20 amino acids. Amino acid deficiency ciated with their metaboism.
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, TABLE 27–2 Enzymes Required for the Synthesis of
which resuts when a chid is weaned onto a starchy diet poor in Amino Acids From Amphibolic Intermediates
protein, and marasmus, in which both caoric intake and spe-
cific amino acids are deficient. 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- Synthetase, & Aminotransferases
genase (EC 1.4.1.3). 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,
BIOSYNTHESIS OF THE 27–2, and 27–4) resuts in the incorporation of potentiay cyto-
NUTRITIONALLY NONESSENTIAL toxic ammonium ion into nontoxic amino acids.
AMINO ACIDS
Asparagine
Glutamate The conversion of aspartate to asparagine, catayzed by aspara-
Gutamate, the precursor of the so-caed “gutamate famiy” of gine synthetase (Figure 27–5), resembes the gutamine syn-
amino acids, is formed by the reductive amidation of the citric thetase reaction (see Figure 27–2), but gutamine, rather than
acid cyce intermediate α-ketogutarate, a reaction catayzed ammonium ion, provides the nitrogen. Bacteria asparagine
by mitochondria gutamate dehydrogenase (Figure 27–1). synthetases can, however, aso use ammonium ion. The reac-
The reaction both strongy favors gutamate synthesis and tion invoves the intermediate formation of asparty phosphate
owers the concentration of cytotoxic ammonium ion. (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.
Glutamine
The amidation of gutamate to gutamine catayzed by gutamine Serine
synthetase (Figure 27–2) invoves the intermediate formation of
Oxidation of the α-hydroxy group of the gycoytic interme-
γ-gutamy phosphate (Figure 27–3). Foowing the ordered bind-
diate 3-phosphogycerate, catayzed by 3-phosphogycerate
ing of gutamate and ATP, gutamate attacks the γ-phosphorus of
dehydrogenase, converts it to 3-phosphohydroxypyruvate.
ATP, forming γ-gutamy phosphate and ADP. NH4+ then binds,
Transamination and subsequent dephosphoryation then
and uncharged NH3 attacks γ-gutamy phosphate. Reease of Pi
form serine (Figure 27–7).
and of a proton from the γ-amino group of the tetrahedra inter-
mediate then aows reease of the product, gutamine.
Glycine
Gycine aminotransferases can catayze the synthesis of gy-
Alanine & Aspartate cine from gyoxyate and gutamate or aanine. Unike most
Transamination of pyruvate forms aanine (Figure 27–4). Simi- aminotransferase reactions, is strongy favored gycine syn-
ary, transamination of oxaoacetate forms aspartate. 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 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 27–10 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 FIGURE 27–12 Conversion of phenylalanine to tyrosine by
deficiency of the vitamin C required for these two hydroxyases phenylalanine hydroxylase (EC 1.14.16.1). Two distinct enzymatic
activities are involved. Activity II catalyzes reduction of dihydrobiop-
resuts in scurvy, in which beeding gums, sweing joints, and terin by NADPH, and activity I the reduction of O2 to H2O and of phe-
impaired wound heaing resut from the impaired stabiity of co- nylalanine to tyrosine. This reaction is associated with several defects
agen (see Chapters 5 and 50). of phenylalanine metabolism discussed in Chapter 29.
278 SECTION VI Metabolism of Proteins & Amino Acids

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
FIGURE 27–13 Hydroxylation of a proline-rich peptide. can then be incorporated into proteins.
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) SUMMARY
catalyzes an analogous reaction. ■ A vertebrates can form certain amino acids from amphiboic
intermediates or from other dietary amino acids. The
These α-keto acids thus can repace their corresponding amino intermediates and the amino acids to which they give rise are
acids in the diet. α-ketogutarate (Gu, Gn, Pro, Hyp), oxaoacetate (Asp, Asn),
and 3-phosphogycerate (Ser, Gy).
Selenocysteine, the 21st Amino Acid ■ Cysteine, tyrosine, and hydroxyysine are formed from
nutritionay essentia amino acids. Serine provides the carbon
Whie the occurrence of seenocysteine (Figure 27–14) in pro- skeeton and homocysteine the sufur for cysteine biosynthesis.
teins is reativey uncommon, at east 25 human seenoproteins ■ In scurvy, a nutritiona disease that resuts from a deficiency
are known. Seenocysteine is present at the active site of severa of vitamin C, impaired hydroxyation of peptidy proine and
human enzymes that catayze redox reactions. Exampes incude peptidy ysine resuts in a faiure to provide the substrates for
thioredoxin reductase, gutathione peroxidase, and the deiodinase cross-inking of maturing coagens.
that converts thyroxine to triiodothyronine. Where present, ■ Phenyaanine hydroxyase converts phenyaanine to tyrosine.
seenocysteine participates in the cataytic mechanism of these Since the reaction catayzed by this mixed function oxidase is
enzymes. Significanty, the repacement of seenocysteine by irreversibe, tyrosine cannot give rise to phenyaanine.
cysteine can actuay reduce cataytic activity. Impairments in ■ Neither dietary hydroxyproine nor hydroxyysine is
human seenoproteins have been impicated in tumorgenesis incorporated into proteins because no codon or tRNA dictates
and atheroscerosis, and are associated with seenium deficiency their insertion into peptides.
cardiomyopathy (Keshan disease). ■ Peptidy hydroxyproine and hydroxyysine are formed
Biosynthesis of seenocysteine requires serine, seenate by hydroxyation of peptidy proine or ysine in reactions
(SeO42−), ATP, a specific tRNA, and severa enzymes. Serine pro- catayzed by mixed-function oxidases that require vitamin C as
vides the carbon skeeton of seenocysteine. Seenophosphate, cofactor.
formed from ATP and seenate (see Figure 27–14), serves as the ■ Seenocysteine, an essentia active site residue in severa
seenium donor. Unike 4-hydroxyproine or 5-hydroxyysine, mammaian enzymes, arises by cotransationa insertion from a
seenocysteine arises cotranslationally during its incorporation previousy modified tRNA.
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 REFERENCES
codon invoves the seenocysteine insertion eement, a stem- Gadyshev VN, Arner ES, Brigeius FR, et a: Seenoprotein gene
oop structure in the untransated region of the mRNA. tRNASec nomencature. J Bio Chem 2016;291:20436.
is first charged with serine by the igase that charges tRNASer. Kiberg MS: Asparagine synthetase chemotherapy. Annu Rev
Biochem 2006;75:629.
Rayman RP: Seenium and human heath. Lancet
H
2012;379;9822;1256.
H Se CH2 C COO– Ruzzo EK, Capo-Chichi JM, Ben-Zeev B, et a: Deficiency of
NH3+ asparagine synthetase causes congenita microcephay and a
O
progressive form of encephaopathy. Neuron 2013;80:429.
Sticke F, Inderbitzin D, Candinas D: Roe of nutrition in iver
Se + ATP + H2O AMP + Pi + H Se P O– transpantation for end-stage chronic iver disease. Nutr Rev
O – 2008;66:47.
Turanov AA, Shchedrina VA, Everey RA, et a: Seenoprotein S
FIGURE 27–14 Selenocysteine (top) and the reaction cata- is invoved in maintenance and transport of mutiprotein
lyzed by selenophosphate synthetase (EC 2.7.9.3) (bottom). compexes. Biochem J 2014;462:555.
 C H A P T E R

Catabolism o Proteins &

o Amino Acid Nitrogen
Victor W. Rodwell, PhD
28
OBJ E C TI VE S ■ Describe protein turnover, indicate the mean rate o protein turnover in healthy
individuals, and provide examples o human proteins that are degraded at rates
After studying this chapter, greater than the mean rate.
you should be able to: ■ Outline the events in protein turnover by both ATP-dependent and ATP-
independent pathways, and indicate the roles in protein degradation
played by the proteasome, ubiquitin, cell surace receptors, circulating
asialoglycoproteins, and lysosomes.
■ Indicate how the ultimate end products o nitrogen catabolism in mammals
dier rom those in birds and fsh.
■ Illustrate the central roles o transaminases (aminotranserases), o glutamate
dehydrogenase, and o glutaminase in human nitrogen metabolism.
■ Use structural ormulas to represent the reactions that convert NH3, CO2, and
the amide nitrogen o aspartate into urea, and identiy the subcellular locations
o the enzymes that catalyze urea biosynthesis.
■ Indicate the roles o allosteric regulation and o acetylglutamate in the
regulation o the earliest steps in urea biosynthesis.
■ Explain why metabolic deects in dierent enzymes o urea biosynthesis,
although distinct at the molecular level, present similar clinical signs and
symptoms.
■ Describe both the classical approaches and the role o tandem mass
spectrometry in screening neonates or inherited metabolic diseases.

BIOMEDICAL IMPORTANCE o amino acids. issues thereore convert ammonia to the

amide nitrogen o the nontoxic amino acid gutamine. Subse-
In norma aduts, nitrogen intake matches nitrogen excreted. quent deamination o gutamine in the iver reeases ammonia,
Positive nitrogen baance, an excess o ingested over excreted which is eicienty converted to urea, which is not toxic. How-
nitrogen, accompanies growth and pregnancy. Negative nitro- ever, i iver unction is compromised, as in cirrhosis or hepa-
gen baance, where output exceeds intake, may oow surgery, titis, eevated bood ammonia eves generate cinica signs and
advanced cancer, and the nutritiona disorders kwashiorkor symptoms. Each enzyme o the urea cyce provides exampes
and marasmus. Genetic disorders that resut rom deects in o metaboic deects and their physioogic consequences. In
the genes that encode ubiquitin, ubiquitin igases, or deubiq- addition, the urea cyce provides a useu moecuar mode or
uitinating enzymes that participate in the degradation o cer- the study o other human metaboic deects.
tain proteins incude Angeman syndrome, juvenie Parkinson
disease, von Hippe-Lindau syndrome, and congenita poy-
cythemia. his chapter describes how the nitrogen o amino PROTEIN TURNOVER
acids is converted to urea, and the metaboic disorders that he continuous degradation and synthesis (turnover) o ce-
accompany deects in this process. Ammonia, which is highy uar proteins occur in a orms o ie. Each day, humans turn
toxic, arises in humans primariy rom the α-amino nitrogen over 1 to 2% o their tota body protein, principay musce

279
280 SECTION VI Metabolism o Proteins & Amino Acids

protein. High rates o protein degradation occur in tissues

N-terminus
that are undergoing structura rearrangement, or exampe,
uterine tissue during pregnancy, skeeta musce in starvation, Lysine 63
and tadpoe tai tissue during metamorphosis. Whie approxi-
matey 75% o the amino acids iberated by protein degrada-
tion are reutiized, the remaining excess ree amino acids are
not stored or uture use. Amino acids not immediatey incor-
porated into new protein are rapidy degraded. he major por-
C-terminus
tion o the carbon skeetons o the amino acids is converted to
Lysine 48
amphiboic intermediates, whie in humans the amino nitro-
gen is converted to urea and excreted in the urine. FIGURE 28–1 Three-dimensional structure of ubiquitin.
Shown are α-helices (blue), β-strands (green), and the R-groups
o lysyl residues (orange). Lys48 & Lys63 are sites or attachment
PROTEASES & PEPTIDASES o additional ubiquitin molecules during polyubiquitination.
(Rogerdodd/Wikipedia)
DEGRADE PROTEINS TO
AMINO ACIDS
he reative susceptibiity o a protein to degradation is expressed avor ubiquitination. Attachment o a singe ubiquitin moecue
as its haf-ife (t1/2), the time required to ower its concentration to transmembrane proteins aters their subceuar ocaization
to ha o its initia vaue. Ha-ives o iver proteins range rom and targets them or degradation. Soube proteins undergo
under 30 minutes to over 150 hours. ypica “housekeeping” poyubiquitination, the igase-catayzed attachment o our or
enzymes such as those o gycoysis, have t1/2 vaues o over more additiona ubiquitin moecues (Figure 28–1). Subsequent
100 hours. By contrast, key reguatory enzymes may have t1/2 degradation o ubiquitin-tagged proteins takes pace in the
vaues as ow as 0.5 to 2 hours. PES sequences, regions rich in proteasome, a macromoecue that aso is ubiquitous in
proine (P), gutamate (E), serine (S), and threonine (), target
some proteins or rapid degradation. Intraceuar proteases
hydroyze interna peptide bonds. he resuting peptides are
then degraded to amino acids by endopeptidases that hydro-
yze interna peptide bonds, and by aminopeptidases and
carboxypeptidases that remove amino acids sequentiay rom
the amino- and carboxy-termini, respectivey.

ATP-Independent Degradation
Degradation o bood gycoproteins (see Chapter 46) oows
oss o a siaic acid moiety rom the nonreducing ends o their
oigosaccharide chains. Asiaogycoproteins are then interna-
ized by iver-ce asiaogycoprotein receptors and degraded
by ysosoma proteases. Extraceuar, membrane-associated,
and ong-ived intraceuar proteins are aso degraded in yso-
somes by AP-independent processes.

ATP & Ubiquitin-Dependent

Degradation
Degradation o reguatory proteins with short ha-ives and
o abnorma or misoded proteins occurs in the cytoso, and
requires AP and ubiquitin. Named based on its presence in a
eukaryotic ces, ubiquitin is a sma (8.5 kDa, 76 residue) poypep-
tide that targets many intraceuar proteins or degradation. he
FIGURE 28–2 Reactions involved in the attachment of
primary structure o ubiquitin is highy conserved. Ony 3 o 76 ubiquitin (Ub) to proteins. Three enzymes are involved. E1 is an acti-
residues dier between yeast and human ubiquitin. Figure 28–1 vating enzyme, E2 a transerase, and E3 a ligase. While depicted as
iustrates the three-dimensiona structure o ubiquitin. Ubiquitin single entities, there are several types o E1, and over 500 types o E2.
moecues are attached by non–α-peptide bonds ormed between The terminal COOH o ubiquitin irst orms a thioester. The coupled
hydrolysis o PPi by pyrophosphatase ensures that the reaction will
the carboxy termina o ubiquitin and the ε-amino groups o ysy
proceed readily. A thioester exchange reaction now transers acti-
residues in the target protein (Figure 28–2). he residue present vated ubiquitin to E2. E3 then catalyzes the transer o ubiquitin to
at its amino terminus aects whether a protein is ubiquitinated. the ε-amino group o a lysyl residue o the target protein. Additional
Amino termina Met or Ser residues retard, whereas Asp or Arg rounds o ubiquitination result in subsequent polyubiquitination.
 CHAPTER 28 Catabolism o Proteins & o Amino Acid Nitrogen 281

Ub
Ub
Ub
Ub

Regulatory
particle

Gated
pore

Core particle

Active
sites

FIGURE 28–4 An end-on view of a proteasome.

Gated (Thomas Splettstoesser/Wikipedia)
Regulatory
pore
particle

utiization by various tissues. Musce generates over ha o the

tota body poo o ree amino acids, and iver is the site o the
FIGURE 28–3 Representation of the structure of a protea- urea cyce enzymes necessary or disposa o excess nitrogen.
some. The upper ring is gated to permit only polyubiquitinated pro-
Musce and iver thus pay major roes in maintaining circuat-
teins to enter the proteosome, where immobilized internal proteases
degrade them to peptides. ing amino acid eves.
Figure 28–5 summarizes the postabsorptive state. Free
amino acids, particuary aanine and gutamine, are reeased
eukaryotic ces. he proteasome consists o a macromoecuar, rom musce into the circuation. Aanine is extracted primar-
cyindrica compex o proteins, whose stacked rings orm a cen- iy by the iver, and gutamine is extracted by the gut and the
tra pore that harbors the active sites o proteoytic enzymes. For kidney, both o which convert a signiicant portion to aanine.
degradation, a protein thus must irst enter the centra pore. Entry Gutamine aso serves as a source o ammonia or excretion
into the core is reguated by the two outer rings that recognize by the kidney. he kidney provides a major source o serine
poyubiquitinated proteins (Figures 28–3 and 28–4). or uptake by periphera tissues, incuding iver and musce.
For the discovery o ubiquitin-mediated protein degradation,
Aaron Ciechanover and Avram Hershko o Israe and Irwin
Rose o the United States were awarded the 2004 Nobe Prize in Kidney
Chemistry. Genetic disorders that resut rom deects in the genes NH3
that encode ubiquitin, ubiquitin igases, or deubiquitinating Brain
enzymes incude Angeman syndrome, autosoma recessive
juvenie Parkinson disease, von Hippe-Lindau syndrome, and
congenita poycythemia. For additiona aspects o protein
degradation and o ubiquitination, incuding its roe in the ce Val Ser
cyce, see Chapters 4 and 35. Gut Ala
Gln
Ala

INTERORGAN EXCHANGE Urea

MAINTAINS CIRCULATING LEVELS Ala Glucose

OF AMINO ACIDS Muscle

Liver

he maintenance o steady-state concentrations o circuat- FIGURE 28–5 Interorgan amino acid exchange in normal
ing pasma amino acids between meas depends on the net postabsorptive humans. The key role o alanine in amino acid out-
baance between reease rom endogenous protein stores and put rom muscle and gut and uptake by the liver is shown.
282 SECTION VI Metabolism o Proteins & Amino Acids

state, they provide the brain with an energy source, and post-
Liver Blood Muscle
prandiay they are extracted predominanty by musce, having
Glucose
been spared by the iver.

ANIMALS CONVERT α-AMINO

Glucose
Glucose NITROGEN TO VARIED END
Urea PRODUCTS
Pyruvate Pyruvate
– NH2 Depending on their ecoogica niche and physioogy, dier-
–NH2 ent animas excrete excess nitrogen as ammonia, uric acid, or
Alanine Alanine urea. he aqueous environment o teeostean ish, which are
Amino acids
ammonoteic (excrete ammonia), permits them to excrete
Alanine water continuousy to aciitate excretion o ammonia, which
is highy toxic. Whie this approach is appropriate or an
aquatic anima, birds must both conserve water and maintain
ow weight. Birds, which are uricoteic, address both prob-
ems by excreting nitrogen-rich uric acid (see Figure 33–11)
as semisoid guano. Many and animas, incuding humans,
FIGURE 28–6 The glucose-alanine cycle. Alanine is synthe- are ureoteic and excrete nontoxic, highy water-soube urea.
sized in muscle by transamination o glucose-derived pyruvate, Since urea is nontoxic to humans, high bood eves in rena
released into the bloodstream, and taken up by the liver. In the
liver, the carbon skeleton o alanine is reconverted to glucose and disease are a consequence, not a cause, o impaired rena
released into the bloodstream, where it is available or uptake by unction.
muscle and resynthesis o alanine.

BIOSYNTHESIS OF UREA
Branched-chain amino acids, particuary vaine, are reeased Urea biosynthesis occurs in our stages: (1) transamination,
by musce and taken up predominanty by the brain. (2) oxidative deamination o gutamate, (3) ammonia trans-
Aanine is a key guconeogenic amino acid (Figure 28–6). port, and (4) reactions o the urea cyce (Figure 28–8). he
he rate o hepatic guconeogenesis rom aanine is ar higher expression in iver o the RNAs or a the enzymes o the urea
than rom a other amino acids. he capacity o the iver or cyce increases severaod in starvation, probaby secondary
guconeogenesis rom aanine does not reach saturation unti to enhanced protein degradation to provide energy.
the aanine concentration reaches 20 to 30 times its norma
physioogic eve. Foowing a protein-rich mea, the spanchnic
tissues reease amino acids (Figure 28–7) whie the periphera
Transamination Transfers α-Amino
musces extract amino acids, in both instances predominanty Nitrogen to α-Ketoglutarate,
branched-chain amino acids. Branched-chain amino acids Forming Glutamate
thus serve a specia roe in nitrogen metaboism. In the asting ransamination reactions interconvert pairs o α-amino
acids and α-keto acids (Figure 28–9). ransamination reac-
Kidney tions, which are reey reversibe, aso unction in amino acid

Brain

Ala
(20
%
Val amBran
Po ino ch
Gln Gut r ta ac ed-
lc ids ch
irc ) ain
ula
tio
n

(60% Branched-chain amino acids)

Muscle Liver
Ala

FIGURE 28–7 Summary of amino acid exchange between FIGURE 28–8 Overall flow of nitrogen in amino acid
organs immediately after feeding. catabolism.
 CHAPTER 28 Catabolism o Proteins & o Amino Acid Nitrogen 283

NH +3 O R COO–
CH
CH O– C O–
R1 C R1 C
N
O O

HO CH2
OPO3–2
O NH + 3
H3C N
C O– CH O–
R2 C R2 C

O O FIGURE 28–11 Structure of a Schiff base formed between

pyridoxal phosphate and an amino acid.
FIGURE 28–9 Transamination. The reaction is reely reversible
with an equilibrium constant close to unity.
associated with eevated serum eves o aminotranserases
(see abe 7–1).
biosynthesis (see Figure 27–4). A o the common amino acids
except ysine, threonine, proine, and hydroxyproine partici-
pate in transamination. ransamination is not restricted to l-GLUTAMATE DEHYDROGENASE
α-amino groups. he δ-amino group o ornithine (but not the OCCUPIES A CENTRAL POSITION
ε-amino group o ysine) readiy undergoes transamination.
Aanine-pyruvate aminotranserase (aanine aminotrans-
IN NITROGEN METABOLISM
erase, EC 2.6.1.2) and gutamate-α-ketogutarate aminotrans- ranser o amino nitrogen to α-ketogutarate orms l-gutamate.
erase (gutamate aminotranserase, EC 2.6.1.1) catayze the Hepatic l-gutamate dehydrogenase (GDH), which can use
transer o amino groups to pyruvate (orming aanine) or to either NAD+ or NADP+, reeases this nitrogen as ammonia
α-ketogutarate (orming gutamate). (Figure 28–12). Conversion o α-amino nitrogen to ammo-
Each aminotranserase is speciic or one pair o substrates, nia by the concerted action o gutamate aminotranserase and
but nonspeciic or the other pair. Since aanine is aso a sub- GDH is oten termed “transdeamination.” Liver GDH activ-
strate or gutamate aminotranserase, the α-amino nitrogen ity is aostericay inhibited by AP, GP, and NADH, and is
rom a amino acids that undergo transamination can be con- activated by ADP. he GDH reaction is reey reversibe, and
centrated in gutamate. his is important because l-gutamate aso unctions in amino acid biosynthesis (see Figure 27–1).
is the ony amino acid that undergoes oxidative deamination
at an appreciabe rate in mammaian tissues. he ormation
o ammonia rom α-amino groups thus occurs mainy via the AMINO ACID OXIDASES REMOVE
α-amino nitrogen o l-gutamate. NITROGEN AS AMMONIA
ransamination occurs via a “ping-pong” mechanism l-Amino acid oxidase o iver and kidney convert an amino
characterized by the aternate addition o a substrate and acid to an α-imino acid that decomposes to an α-keto acid with
reease o a product (Figure 28–10). Foowing remova o its reease o ammonium ion (Figure 28–13). he reduced avin
α-amino nitrogen by transamination, the remaining carbon is reoxidized by moecuar oxygen, orming hydrogen perox-
“skeeton” o an amino acid is degraded by pathways discussed ide (H2O2), which then is spit to O2 and H2O by cataase, EC
in Chapter 29. 1.11.1.6.
Pyridoxa phosphate (PLP), a derivative o vitamin B6, is
present at the cataytic site o a aminotranserases, and pays
a key roe in cataysis. During transamination, PLP serves as
Ammonia Intoxication Is
a “carrier” o amino groups. An enzyme-bound Schi base Life-Threatening
(Figure 28–11) is ormed between the oxo group o enzyme- he ammonia produced by enteric bacteria and absorbed into
bound PLP and the α-amino group o an α-amino acid. he porta venous bood and the ammonia produced by tissues are
Schi base can rearrange in various ways. In transamination, rapidy removed rom circuation by the iver and converted
rearrangement orms an α-keto acid and enzyme-bound pyri- to urea. hus, normay, ony traces (10-20 μg/dL) are present
doxamine phosphate. As noted earier, certain diseases are in periphera bood. his is essentia, since ammonia is toxic

Pyr Glu
Ala CHO CH2NH2 KG CH2NH2 CHO
E CHO E E E CH2NH2 E E E CHO
Ala Pyr KG Glu

FIGURE 28–10 “Ping-pong” mechanism for transamination. E—CHO and E—CH2NH2 represent enzyme-bound pyridoxal phosphate
and pyridoxamine phosphate, respectively. (Ala, alanine; Glu, glutamate; KG, α-ketoglutarate; Pyr, pyruvate.)
284 SECTION VI Metabolism o Proteins & Amino Acids

NH +
3

–O CH 2 CH O–
C CH 2 C

O O
L-Glutamate
Mg-ATP NH +
4
FIGURE 28–12 The reaction catalyzed by glutamate dehy-
drogenase, EC 1.4.1.2. NAD(P)+ means that either NAD+ or NADP+ Glutamine
synthetase
can serve as the oxidoreductant. The reaction is reversible, but
strongly avors glutamate ormation. Mg-ADP H 2O
+ Pi
NH +
3

H2 N CH 2 CH O–
C CH 2 C
to the centra nervous system. Shoud porta bood bypass the
iver, systemic bood ammonia may reach toxic eves. his O O
occurs in severey impaired hepatic unction or the deveop- L-Glutamine

ment o coatera inks between the porta and systemic veins

in cirrhosis. Symptoms o ammonia intoxication incude
FIGURE 28–14 Formation of glutamine, catalyzed by gluta-
mine synthetase, EC 6.3.1.2.
tremor, surred speech, burred vision, coma, and utimatey
death. Ammonia may be toxic to the brain in part because it
reacts with α-ketogutarate to orm gutamate. he resuting deiciency in neonate gutamine synthetase resuts in severe
depetion o α-ketogutarate then impairs unction o the tri- brain damage, mutiorgan aiure, and death.
carboxyic acid (CA) cyce in neurons.
Glutaminase & Asparaginase
Glutamine Synthetase Fixes Ammonia Deamidate Glutamine & Asparagine
as Glutamine here are two human isoorms o mitochondria gutamin-
ase, termed iver-type and rena-type gutaminase. Products
Formation o gutamine is catayzed by mitochondria gutamine
o dierent genes, the gutaminases dier with respect to their
synthetase (Figure 28–14). Since amide bond synthesis is
structure, kinetics, and reguation. Hepatic gutaminase eves
couped to the hydroysis o AP to ADP and Pi, the reac-
rise in response to high protein intake whie rena kidney-type
tion strongy avors gutamine synthesis. During cataysis,
gutaminase increases in metaboic acidosis. Hydroytic reease
gutamate attacks the γ-phosphory group o AP, orming
o the amide nitrogen o gutamine as ammonia, catayzed by
γ-gutamy phosphate and ADP. Foowing deprotonation o
gutaminase (Figure 28–15), strongy avors gutamate orma-
NH4+, NH3 attacks γ-gutamy phosphate, and gutamine and
tion. An anaogous reaction is catayzed by l-asparaginase
Pi are reeased. In addition to providing gutamine to serve
(EC 3.5.1.1). he concerted action o gutamine synthetase
as a carrier o nitrogen, carbon and energy between organs
and gutaminase thus catayzes the interconversion o ree
(Figure 28–5), gutamine synthetase pays a major roe both in
ammonium ion and gutamine.
ammonia detoxiication and in acid–base homeostasis. A rare
NH +
3

H2 N CH 2 CH O–
C CH 2 C

O O
L-Glutamine
H2O

Glutaminase

NH +
4

NH +
3

–O CH 2 CH O–
C CH 2 C

O O
L-Glutamate

FIGURE 28–15 The reaction catalyzed by glutaminase, EC

FIGURE 28–13 Oxidative deamination catalyzed by l-amino 3.5.1.2. The reaction proceeds essentially irreversibly in the direction
acid oxidase (l-α-amino acid:O2 oxidoreductase, EC 1.4.3.2). The o glutamate and NH4+ ormation. Note that the amide nitrogen, not
α-imino acid, shown in brackets, is not a stable intermediate. the α-amino nitrogen, is removed.
 CHAPTER 28 Catabolism o Proteins & o Amino Acid Nitrogen 285

Formation & Secretion of Ammonia others serve as carriers o the atoms that utimatey become urea.
he major metaboic roe o ornithine, citruine, and arginino-
Maintains Acid–Base Balance succinate in mammas is urea synthesis. Urea synthesis is a cycic
Excretion into urine o ammonia produced by rena tubuar process. Whie ammonium ion, CO2, AP, and aspartate are con-
ces aciitates cation conservation and reguation o acid– sumed, the ornithine consumed in reaction 2 is regenerated in
base baance. Ammonia production rom intraceuar rena reaction 5. hus, there is no net oss or gain o ornithine, citru-
amino acids, especiay gutamine, increases in metaboic aci- ine, argininosuccinate, or arginine. As indicated in Figure 28–16,
dosis and decreases in metaboic akaosis. some reactions o urea synthesis occur in the matrix o the mito-
chondrion, and other reactions in the cytoso.
Urea Is the Major End Product of
Nitrogen Catabolism in Humans Carbamoyl Phosphate Synthetase I
Synthesis o 1 mo o urea requires 3 mo o AP, 1 mo each Initiates Urea Biosynthesis
o ammonium ion and o aspartate, and empoys ive enzymes Condensation o CO2, ammonia, and AP to orm carbamoy
(Figure 28–16). O the six participating amino acids, phosphate is catayzed by mitochondria carbamoy phos-
N-acetygutamate unctions soey as an enzyme activator. he phate synthetase I (EC 6.3.4.16). A cytosoic orm o this

FIGURE 28–16 Reactions and intermediates of urea biosynthesis. The nitrogen-containing groups that contribute to the ormation o
urea are shaded. Reactions 1 and 2 occur in the matrix o liver mitochondria and reactions 3 , 4 , and 5 in liver cytosol. CO2 (as bicarbon-
ate), ammonium ion, ornithine, and citrulline enter the mitochondrial matrix via speciic carriers (see red dots) present in the inner membrane
o liver mitochondria.
286 SECTION VI Metabolism o Proteins & Amino Acids

enzyme, carbamoy phosphate synthetase II, uses gutamine aminotranserase then reorms aspartate. he carbon skeeton
rather than ammonia as the nitrogen donor and unctions o aspartate-umarate thus acts as a carrier o the nitrogen o
in pyrimidine biosynthesis (see Figure 33–9). he concerted gutamate into a precursor o urea.
action o gutamate dehydrogenase and carbamoy phosphate
synthetase I thus shuttes amino nitrogen into carbamoy
phosphate, a compound with high group transer potentia.
Cleavage of Arginine Releases Urea &
Carbamoy phosphate synthetase I, the rate-imiting Reforms Ornithine
enzyme o the urea cyce, is active ony in the presence o Hydroytic ceavage o the guanidino group o arginine, cata-
N-acetygutamate, an aosteric activator that enhances the yzed by iver arginase (EC 3.5.3.1), reeases urea (reaction
ainity o the synthetase or AP. Synthesis o 1 mo o car- 5, Figure 28–16). he other product, ornithine, reenters iver
bamoy phosphate requires 2 mo o AP. One AP serves as mitochondria and participates in additiona rounds o urea
the phosphory donor or ormation o the mixed acid anhy- synthesis. Ornithine and ysine are potent inhibitors o argi-
dride bond o carbamoy phosphate. he second AP provides nase, and compete with arginine. Arginine aso serves as the
the driving orce or synthesis o the amide bond o carbamoy precursor o the potent musce reaxant nitric oxide (NO) in a
phosphate. he other products are 2 mo o ADP and 1 mo o Ca2+-dependent reaction catayzed by NO synthetase.
Pi (reaction 1, Figure 28–16). he reaction proceeds stepwise.
Reaction o bicarbonate with AP orms carbony phosphate
and ADP. Ammonia then dispaces ADP, orming carbamate
Carbamoyl Phosphate Synthetase I
and orthophosphate. Phosphoryation o carbamate by the Is the Pacemaker Enzyme of the
second AP then orms carbamoy phosphate. Urea Cycle
he activity o carbamoy phosphate synthetase I is determined
Carbamoyl Phosphate Plus Ornithine by N-acetygutamate, whose steady-state eve is dictated by
Forms Citrulline the baance between its rate o synthesis rom acety-CoA and
l-Ornithine transcarbamoyase (EC 2.1.3.3) catayzes trans- gutamate and its rate o hydroysis to acetate and gutamate,
er o the carbamoy group o carbamoy phosphate to orni- reactions catayzed by N-acetygutamate synthetase (NAGS)
thine, orming citruine and orthophosphate (reaction 2, and N-acetygutamate deacyase (hydroase), respectivey.
Figure 28–16). Whie the reaction occurs in the mitochon- Acety-CoA + l-gutamate → N-acety-l-gutamate + CoASH
dria matrix, both the ormation o ornithine and the subse-
N-acety-l-gutamate + H2O → l-gutamate + acetate
quent metaboism o citruine take pace in the cytoso. Entry
o ornithine into mitochondria and exodus o citruine rom Major changes in diet can increase the concentrations o
mitochondria invoves the mitochondria inner membrane individua urea cyce enzymes 10- to 20-od. For exampe,
carriers ORC1, ORC2, and SLCA25A29 (Figure 28–16). starvation eevates enzyme eves, presumaby to cope with the
increased production o ammonia that accompanies enhanced
Citrulline Plus Aspartate Forms starvation-induced degradation o protein.
Argininosuccinate
Argininosuccinate synthetase (EC 6.3.4.5) inks aspartate GENERAL FEATURES OF
and citruine via the amino group o aspartate (reaction 3,
Figure 28–16), which provides the second nitrogen o urea.
METABOLIC DISORDERS
he reaction requires AP and invoves intermediate orma- he comparativey rare, but we-characterized and medicay
tion o citruy-AMP. Subsequent dispacement o AMP by devastating metaboic disorders associated with the enzymes
aspartate then orms argininosuccinate. o urea biosynthesis iustrate the oowing genera principes
o inherited metaboic diseases:
Cleavage of Argininosuccinate Forms 1. Simiar or identica cinica signs and symptoms can accom-
Arginine & Fumarate pany various genetic mutations in a gene that encodes a
given enzyme or in enzymes that catayze successive reac-
Ceavage o argininosuccinate is catayzed by argininosuccinate
tions in a metaboic pathway.
yase (EC 4.3.2.1). he reaction proceeds with retention o
a three nitrogens in arginine and reease o the aspartate 2. Rationa therapy is based on an understanding o the re-
skeeton as umarate (reaction 4, Figure 28–16). Subsequent evant biochemica enzyme-catayzed reactions in both nor-
addition o water to umarate orms l-maate, whose subse- ma and impaired individuas.
quent NAD+-dependent oxidation orms oxaoacetate. hese 3. he identiication o intermediates and o anciary prod-
two reactions are anaogous to reactions o the citric acid ucts that accumuate prior to a metaboic bock provides
cyce, but are catayzed by cytosolic fumarase and maate the basis or metaboic screening tests that can impicate
dehydrogenase. ransamination o oxaoacetate by gutamate the reaction that is impaired.
 CHAPTER 28 Catabolism o Proteins & o Amino Acid Nitrogen 287

4. Deinitive diagnosis invoves quantitative assay o the activ- suicient protein, arginine, and energy to promote growth
ity o the enzyme suspected to be deective. Assas/test-
and deveopment whie simutaneousy minimizing the meta-
-
5. he DNA sequence o the gene that encodes a given mutant boic perturbations.
enzyme is compared to that o the wid-type gene to iden-
tiy the speciic mutation(s) that cause the disease. Carbamoyl Phosphate Synthetase I
6. he exponentia increase in DNA sequencing o human N-Acetygutamate is essentia or the activity o carbamoy
genes has -identiied dozens o mutations o an aected phosphate synthetase I, EC 6.3.4.16 (reaction 1, Figure 28–16).
gene that are benign or are associated with symptoms o Deects in carbamoy phosphate synthetase I are responsibe
varying severity o a given metaboic disorder. or the reativey rare (estimated requency 1:62,000) meta-
boic disease termed “hyperammonemia type 1.”

METABOLIC DISORDERS ARE N-Acetylglutamate Synthetase

ASSOCIATED WITH EACH N-Acetygutamate synthetase, EC 2.3.1.1 (NAGS), catayzes
REACTION OF THE UREA CYCLE the ormation rom acety-CoA and gutamate o the N-
Five we-documented diseases represent deects in the bio- acetygutamate essentia or carbamoy phosphate synthetase
synthesis o enzymes o the urea cyce. Moecuar genetic I activity.
anaysis has pinpointed the oci o mutations associated with l-Gutamate + acety-CoA → N-acety-l-gutamate + CoASH
each deiciency, each o which exhibits considerabe genetic
and phenotypic variabiity (Tabe 28–1). Whie the cinica and biochemica eatures o NAGS dei-
Urea cyce disorders are characterized by hyperammo- ciency are indistinguishabe rom those arising rom a deect
nemia, encephaopathy, and respiratory akaosis. Four o the
ive metaboic diseases, deiciencies o carbamoy phosphate
O in carbamoy phosphate synthetase I, a deiciency in NAGS
may respond to administered N-acetygutamate.
synthetase I, ornithine carbamoy transerase, argininosuc-
cinate synthetase, and argininosuccinate yase, resut in the Ornithine Permease
accumuation o precursors o urea, principay ammonia and he hyperornithinemia, hyperammonemia, and homocitru-
gutamine. Ammonia intoxication is most severe when the inuria (HHH) syndrome resuts rom mutation o the ORC1
metaboic bock occurs at reactions 1 or 2 (Figure 28–16), or gene that encodes the mitochondria membrane ornithine
i citruine can be synthesized, some ammonia has aready carrier. he inabiity to import cytosoic ornithine into the
been removed by being covaenty inked to an organic mitochondria matrix renders the urea cyce inoperabe, with
metaboite. consequent hyperammonemia, and hyperornithinemia due
Cinica symptoms common to a urea cyce disorders to the accompanying accumuation o cytosoic ornithine. In
incude vomiting, avoidance o high-protein oods, intermit- the absence o its norma acceptor (ornithine), mitochondria
tent ataxia, irritabiity, ethargy, and severe menta retardation. carbamoy phosphate carbamoyates ysine to homocitruine,
he most dramatic cinica presentation occurs in u-term resuting in homocitruinuria.
inants who initiay appear norma, then exhibit progressive
ethargy, hypothermia, and apnea due to high pasma ammo-
nia eves. he cinica eatures and treatment o a ive disor- Ornithine Transcarbamoylase
ders are simiar. Signiicant improvement and minimization he X-chromosome–inked deiciency termed “hyperammo-
o brain damage can accompany a ow-protein diet ingested nemia type 2” reects a deect in ornithine transcarbamoyase
as requent sma meas to avoid sudden increases in bood (reaction 2, Figure 28–16). he mothers aso exhibit hyper-
ammonia eves. he goa o dietary therapy is to provide ammonemia and an aversion to high-protein oods. Leves

TABLE 28–1 Enzymes of Inherited Metabolic Disorders of the Urea Cycle

Enzyme Enzyme Catalog Number OMIMa Reference Figure and Reaction

Carbamoyl-phosphate synthetase 1 6.3.4.16 237300 28-13➀

Ornithine carbamoyl transerase

Argininosuccinate synthetase

Argininosuccinate lyase
Y preme e
2.1.3.3

6.3.4.5

4.3.2.1
311250

215700

608310
28-13➁

28-13➂

28-13➃

Arginase 3.5.3.1 608313 28-13➄

a
Online Mendelian inheritance in man database: ncbi.nlm.nih.gov/omim/
288 SECTION VI Metabolism o Proteins & Amino Acids

o gutamine are eevated in bood, cerebrospina uid, and Can Metabolic Disorders Be Rectified
urine, probaby as a resut o enhanced gutamine synthesis in
response to eevated eves o tissue ammonia.
by Gene or Protein Modification
Despite resuts in anima modes using an adenovira vector to
treat citruinemia, at present gene therapy provides no eec-
Argininosuccinate Synthetase tive soution or human subjects. However, direct CRISPR/
In addition to patients who ack detectabe argininosuccinate Cas9-based modiication o a deective enzyme can restore
synthetase activity (reaction 3, Figure 28–16), 25-od eeva- unctiona enzyme activity o cutured human puripotent
tions in Km or citruine have been reported. In the resuting stem ces.
citruinemia, pasma and cerebrospina uid citruine eves
are eevated, and 1 to 2 g o citruine are excreted daiy.
SUMMARY
Argininosuccinate Lyase ■ Human subjects degrade 1 to 2% o their body protein daiy at
rates that vary widey between proteins and with physioogic
Argininosuccinic aciduria, accompanied by eevated eves state. Key reguatory enzymes oen have short ha-ives.
o argininosuccinate in bood, cerebrospina uid, and urine, ■ Proteins are degraded by both AP-dependent and AP-
is associated with riabe, tuted hair (trichorrhexis nodosa). independent pathways. Ubiquitin targets many intraceuar
Both eary- and ate-onset types are known. he metaboic proteins or degradation. Liver ce surace receptors bind
deect is in argininosuccinate yase (reaction 4, Figure 28–16). and internaize circuating asiaogycoproteins destined or
Diagnosis by the measurement o erythrocyte argininosucci- ysosoma degradation.
nate yase activity can be perormed on umbiica cord bood ■ Poyubiquitinated proteins are degraded by proteases
or amniotic uid ces. on the inner surace o a cyindrica macromoecue,
the proteasome. Entry into the proteasome is gated by a
Arginase donut-shaped protein pore that rejects entry to a but
poyubiquitinated proteins.
Hyperargininemia is an autosoma recessive deect in the gene
■ Fishes excrete highy toxic NH3 directy. Birds convert NH3 to
or arginase (reaction 5, Figure 28–16). Unike other urea cyce
uric acid. Higher vertebrates convert NH3 to urea.
disorders, the irst symptoms o hyperargininemia typicay do
■ ransamination channes amino acid nitrogen into
not appear unti age 2 to 4 years. Bood and cerebrospina uid
gutamate. GDH occupies a centra position in nitrogen
eves o arginine are eevated. he urinary amino acid pattern,
metaboism.
which resembes that o ysine-cystinuria (see Chapter 29),
■ Gutamine synthetase converts NH3 to nontoxic gutamine.
may reect competition by arginine with ysine and cysteine
Gutaminase reeases NH3 or use in urea synthesis.
or reabsorption in the rena tubue.
■ NH3, CO2, and the amide nitrogen o aspartate provide the
atoms o urea.
Analysis of Neonate Blood by Tandem ■ Hepatic urea synthesis takes pace in part in the mitochondria
Mass Spectrometry Can Detect matrix and in part in the cytoso.
Metabolic Diseases ■ Changes in enzyme eves and aosteric reguation o
Metaboic diseases caused by the absence or unctiona impair- carbamoy phosphate synthetase I by N-acetygutamate
reguate urea biosynthesis.
ment o metaboic enzymes can be devastating. Eary dietary
intervention, however, can in many instances ameiorate the ■ Metaboic diseases are associated with deects in each enzyme
otherwise inevitabe dire eects. he eary detection o such o the urea cyce, o the ORC1 ornithine carrier, and o NAGS.
metaboic diseases is thus is o primary importance. Since ■ Te metaboic disorders o urea biosynthesis iustrate six
the initiation in the United States o newborn screening pro- genera principes o a metaboic disorders.
grams in the 1960s, a states now conduct metaboic screen- ■ andem mass spectrometry is the technique o choice or
ing o newborn inants. he poweru and sensitive technique screening neonates or inherited metaboic diseases.
o tandem mass spectrometry (MS) (see Chapter 4) can in
a ew minutes detect over 40 anaytes o signiicance in the
detection o metaboic disorders. Most states empoy tandem
REFERENCES
MS to screen newborns to detect metaboic disorders such as Adam S, Ameida MF, Assoun M, et a: Dietary management o
urea cyce disorders: European practice. Mo Genet Metab
organic acidemias, aminoacidemias, disorders o atty acid
2013;110:439.
oxidation, and deects in the enzymes o the urea cyce. An Burgard P, Köker S, Haege G, et a. Neonata mortaity and outcome
artice in Clinical Chemistry 2006 39:315 reviews the theory o at the end o the frst year o ie in eary onset urea cyce
tandem MS, its appication to the detection o metaboic disor- disorders. J Inherit Metab Dis. 2016;39:219.
ders, and situations that can yied ase positives, and incudes Dwane L, Gaagher WM, Ni Chonghaie , et a: Te emerging roe
a engthy tabe o detectabe anaytes and the reevant meta- o non-traditiona ubiquitination in oncogenic pathways. J Bio
boic diseases. Chem 2017;292:3543.