104 9.

Translation

C. Consequences of altering the nucleotide sequence

u Changing a single nucleotide base on the mRNA chain (a "point muta-
(Termination codon)
tion") can lead to any one of three results (Figure 9.3):
Nonsense
1. Silent mutation: The codon containing the changed base may
1mutation
code for the same amino acid. For example, if the serine codon
UCA is given a different third base "U" to become UCU, it still codes
Silent for serine. This is termed a "silent" mutation.
mutation
(Codon for serine)
2. Missense mutation: The codon containing the changed base
Missense
may code for a different amino acid. For example, if the serine
]
mutation
uc ro codon UCA is given a different first base "C" to become CCA, it will
(Codon for serine) code for a different amino acid, in this case, proline. The substitu-
(i CA tion of an incorrect amino acid is called a "missense" mutation.
(Codon for proline)
3. Nonsense mutation: The codon containing the changed base
Figure 9.3 may become a termination codon. For example, if the serine codon
Possible effects of changing a single
UCA is given a different second base "A" to become UAA, the new
nucleotide base in the coding region codon causes termination of translation at that point and the pro-
of the mRNA chain. duction of a shortened (truncated) protein. The creation of a ter-
mination codon at an inappropriate place is called a "nonsense"
mutation.

4. Other mutations: These can alter the amount or structure of the

Huntington disease
protein produced by translation.
Huntington disease
a. Trinucleotide repeat expansion: Occasionally, a sequence of
three bases that is repeated in tandem will become amplified in
number so that too many copies of the triplet occur. If this occurs
within the coding region of a gene, the protein will contain many
Tandem repeats of CAG triplets extra copies of one amino acid. For example, amplification of
coding for glutamine
the CAG codon leads to the insertion of many extra glutamine
mRNA is translated
into huntington residues in the Huntington protein, causing the neurodegenera-

jprotein with
abnormal glutamine
repeats
tive disorder, Huntington disease (Figure 9.4). The additional
glutamines result in unstable proteins that cause the accumu-
lation of protein aggregates. If the trinucleotide repeat expan-
sion occurs in the untranslated portion of a gene, the result can
be a decrease in the amount of protein produced as seen, for
example, in fragile X syndrome and myotonic dystrophy.
Aggregated proteins
and peptides b. Splice site mutations: Mutations at splice sites can alter the
way in which introns are removed from the pre-mRNA mole-
Other triplet expansion diseases cules, producing aberrant proteins.

Fraglle-X syndrome c. Frame-shift mutations: If one or two nucleotides are either

5' AAAA deleted from or added to the coding region of a message
/ ~-~~~'!!!!'!'!, ' sequence, a frame-shift mutation occurs and the reading frame
(CGGh-so
is altered. This can result in a product with a radically different
amino acid sequence (Figure 9.5) or a truncated product due
Myotonic dystrophy to the creation of a termination codon. If three nucleotides are
5' AAAA added, a new amino acid is added to the peptide, or if three
~~~~~~__, /,
(CUG)5-35 nucleotides are deleted, an amino acid is lost. In these instances,
the reading frame is not affected. Loss of three nucleotides main-
Figure 9.4 tains the reading frame but can result in serious pathology. For
Role of tandem triplet repeats in example, cystic fibrosis (CF), a hereditary disease that primarily
mRNA causing Huntington disease affects the pulmonary and digestive systems, is most commonly
and other triplet expansion diseases. caused by a deletion of three nucleotides from the coding region
Ill. Components Required for Translation 105

of a gene, resulting in the loss of phenylalanine at the 508th posi-

( Addition of base )
tion (.!lF508) in the protein encoded by that gene. This .!lF508
mutation prevents normal folding of the CF transmembrane con- mRNA
ductance regulator (CFTR) protein, leading to its destruction by AAA UCA m ccUAUGGCU AAA
L______j ~ L________j L________j

the proteosome (see Chapter 12). CFTR normally functions as Ser Ser Tyr Gly
a chloride channel in epithelial cells, and its loss results in the
production of thick, sticky secretions in the lungs and pancreas,
leading to lung damage and digestive deficiencies. In over 70% of
1!1 1 AddOkm of U

patients with CF, the .!lF508 mutation is the cause of the disease. /VV\ UCA C CUAUGGCU /VV\.
. _ J ' - - - ! ~-..~ ' -
5'- end l .Ser 3' d
Pro Met Ala - en

~
Ill. COMPONENTS REQUIRED FOR TRANSLATION
c """"'" of c
A large number of components are required for the synthesis of a protein.
These include all the amino acids that are found in the finished product, /VV\. U C A C U AUG G C U /VV\
I ..I ~L..__I

the mRNA to be translated, transfer RNA (tRNA), functional ribosomes, Ser Leu Trp
energy sources, and enzymes, as well as protein factors needed for ini-
tiation, elongation, and termination of the polypeptide chain. ll Deletion of base J
A. Amino acids
Figure 9.5
All the amino acids that eventually appear in the finished protein must Frame-shift mutations as a result of
be present at the time of protein synthesis. (Note: If one amino acid addition or deletion of a base can cause
an alteration in the reading frame of
is missing [e.g., if the diet does not contain an essential amino acid],
mRNA.
translation stops at the codon specifying that amino acid. This dem-
onstrates the importance of having all the essential amino acids in
sufficient quantities in the diet to ensure continued protein synthesis.)

B. Transfer RNA Methionine,

ACC
tRNAs are able to carry a specific amino acid and to recognize the 3'
codon for that amino acid. tRNA, therefore, functions as adaptor
5'
molecules.
At least one specific type of tRNA is required per amino acid. In
humans, there are at least 50 species of tRNA, whereas bacteria con-
tain 30 to 40 species. Because there are only 20 different amino acids
commonly carried by tRNA, some amino acids have more than one
specific tRNA molecule. This is particularly true of those amino acids
that are coded for by several codons.

1. Amino acid attachment site: Each tRNA molecule has an attach-

ment site for a specific (cognate) amino acid at its 3' end (Figure 9.6).
The carboxyl group of the amino acid is in an ester linkage with the
3'-hydroxyl group of the ribose moiety of the adenosine nucleotide
in the -CCA sequence at the 3' end of the tRNA. (Note: When a
tRNA has a covalently attached amino acid, it is said to be charged;
when tRNA is not bound to an amino acid, it is described as being
uncharged.) The amino acid that is attached to the tRNA molecule is
said to be activated. Codon

2. Anticodon: Each tRNA molecule also contains a three-base

Figure 9.6
nucleotide sequence-the anticodon-that recognizes a specific
Complementary antiparallel binding of
codon on the mRNA (see Figure 9.6). This codon specifies the the anticodon for methionyl-tRNA (CAU)
insertion into the growing peptide chain of the amino acid carried to the mRNA codon for methionine
by that tRNA. (AUG).
106 9. Translation

Amino acid C. Aminoacyl-tRNA synthetases

Aminoacy/-tRNA
synthetase
~ATP This family of enzymes is required for the attachment of amino acids
to their corresponding tRNA. Each member of this family recognizes
(E) PP;-~>2P; a specific amino acid and the tRNA that corresponds to that amino
acid (isoaccepting tRNA). These enzymes thus implement the genetic
E-AMP-Amino acid
code because they act as molecular dictionaries that can read both
CCA the three-letter code of nucleic acids and the 20-letter code of amino
acids. Each aminoacyl-tRNA synthetase catalyzes a two-step reac-
tion that results in the covalent attachment of the carboxyl group of
an amino acid to the 3' end of its corresponding tRNA. The overall
reaction requires adenosine triphosphate (ATP}, which is cleaved to
adenosine monophosphate (AMP) and inorganic pyrophosphate (PP;)
(Figure 9.7). The extreme specificity of the synthetase in recognizing
both the amino acid and its specific tRNA contributes to the high fidel-
ity of translation of the genetic message. In addition, the synthetases
have a "proofreading" or "editing" activity that can remove mischarged
amino acids from the enzyme or the tRNA molecule.

D. Messenger RNA
The specific mRNA required as a template for the synthesis of the
Aminoacyl-tRNA desired polypeptide chain must be present.

Figure 9.7 E. Functionally competent ribosomes

Attachment of a specific amino acid to Ribosomes are large complexes of protein and ribosomal RNA
its corresponding tRNA by aminoacyl- (rRNA, Figure 9.8).They consist of two subunits-one large and one
tRNA synthetase (E).
small-whose relative sizes are generally given in terms of their sedi-
mentation coefficients, or S (Svedberg) values. (Note: Because the S
values are determined both by shape as well as molecular mass, their
Eukaryotic ribosome numeric values are not strictly additive. The eukaryotic 60S and 40S
subunits form an 80S ribosome.) Prokaryotic and eukaryotic ribo-
somes are similar in structure and serve the same function, namely,
as the "factories" in which the synthesis of proteins occurs.
The larger ribosomal subunit catalyzes the formation of the peptide
bonds that link amino acid residues in a protein. The smaller sub-
unit binds mRNA and is responsible for the accuracy of translation by
ensuring correct base-pairing between the codon in the mRNA and
80S
/ the anticodon of the tRNA.
( 1. Ribosomal RNA: Eukaryotic ribosomes contain four molecules of
I
rRNA (see Figure 9.8). The rRNAs have extensive regions of sec-
ondary structure arising from the base-pairing of the complemen-

~ \~.I
tary sequences of nucleotides in different portions of the molecule.

2. Ribosomal proteins: Ribosomal proteins play a number of roles

55
RNA
~~s; in the structure and function of the ribosome and its interactions
with other components of the translation system.

3. A, P, and E sites on the ribosome: The ribosome has three bind-

285 185
RNA RNA ing sites for tRNA molecules-the A, P, and E sites-each of which
extends over both subunits (see Figure 9.8). Together, they cover
-50 proteins -30 proteins three neighboring codons. During translation, the A site binds
an incoming aminoacyl-tRNA as directed by the codon currently
Figure 9.8 occupying this site. This codon specifies the next amino acid to be
Eukaryotic ribosomal composition. added to the growing peptide chain. The P-site codon is occupied
IV. Codon Recognition by TRNA 107

by peptidyl-tRNA. This tRNA carries the chain of amino acids that

has already been synthesized. The E site is occupied by the empty
tRNA as it is about to exit the ribosome.

4. Cellular location of ribosomes: In eukaryotic cells, the ribo-

somes are either "free" in the cytosol or are in close association
with the endoplasmic reticulum (which is then known as the "rough"
endoplasmic reticulum, or RER). The REA-associated ribosomes
are responsible for synthesizing proteins that are to be exported
from the cell as well as those that are destined to become inte-
grated into plasma, endoplasmic reticulum, or Golgi membranes
or incorporated in lysosomes. Cytosolic ribosomes synthesize pro-
teins required in the cytosol itself or destined for the nucleus, mito-
chondria, and peroxisomes. (Note: Mitochondria contain their own
set of ribosomes and their own unique, circular DNA.) Serine,
ACC
F. Protein factors 3'

Initiation, elongation, and termination (or release) factors are required 5'
for peptide synthesis. Some of these protein factors perform a catalytic
function, whereas others appear to stabilize the synthetic machinery.

G. ATP and GTP are required as sources of energy

Cleavage of four high-energy bonds is required for the addition of
one amino acid to the growing polypeptide chain: two from ATP in
the aminoacyl-tRNA synthetase reaction-one in the removal of PP;
and one in the subsequent hydrolysis of the PP; to inorganic phos-
phate by pyrophosphatase-and two from guanosine triphosphate
(GTP)---one for binding the aminoacyl-tRNA to the A site and one for
the translocation step (see Figure 9.10). (Note: Additional ATP and
GTP molecules are required for initiation in eukaryotes and an addi- mRNA
tional GTP molecule is required for termination.) /VVVVVVVV\. U C G VVVV\/V'VV'\,
s ~

Traditional base- Nontraditional

IV. CODON RECOGNITION BY TRNA pairing observed base-pairing is
in first and second possible between
positions of codon: the third (3')
Correct pairing of the codon in the mRNA with the anticodon of the tRNA position of the
!RNA mRNA codon and the
is essential for accurate translation (see Figure 9.6). Some tRNAs recog-
nize more than one codon for a given amino acid. rr YJ first (5') position
of the anticodon:
[G c)
!RNA mRNA
A. Antiparallel binding between codon and anticodon Iu A) [A u)
[c G)
Binding of the tRNA anticodon to the mRNA codon follows the rules of com-
plementary and antiparallel binding, that is, the mRNA codon is "read" 5' ~
IG 5]
3' by an anticodon pairing in the "flipped" (3' ~ 5') orientation (Figure 9.9). lu ~~
(Note: When writing the sequences of both codons and anticodons, the lc G]
nucleotide sequence must ALWAYS be listed in the 5' ~ 3' order.)

B. Wobble hypothesis D
The mechanism by which tRNAs can recognize more than one
codon for a specific amino acid is described by the "wobble" hypoth- Figure 9.9
esis in which the base at the 5' end of the anticodon (the "first" base Wobble: Nontraditional base-pairing
of the anticodon) is not as spatially defined as the other two bases. between the 5'-nucleotide (first
nucleotide) of the anticodon with the
Movement of that first base allows nontraditional base-pairing with 3'-nucleotide (last nucleotide) of the
the 3' base of the codon (the "last" base of the codon). This move- codon. H, hypoxanthine (the base of
ment is called ''wobble" and allows a single tRNA to recognize more inosine).
108 9. Translation

than one codon. Examples of these flexible pairings are shown in

Figure 9.9. The result of wobbling is that there need not be 61 tRNA
species to read the 61 codons coding for amino acids.

V. STEPS IN PROTEIN TRANSLATION

The pathway of protein synthesis translates the three-letter alpha-

bet of nucleotide sequences on mRNA into the 20-letter alphabet of
amino acids that constitute proteins. The mRNA is translated from its
5' end to its 3' end, producing a protein synthesized from its amino-
terminal end to its carboxyl-terminal end. The process of translation is
divided into three separate steps: initiation, elongation, and termina-
tion. The polypeptide chains produced may be modified by posttrans-
lational modification.

A. Initiation
Initiation of protein synthesis involves the assembly of the compo-
nents of the translation system before peptide bond formation occurs.
These components include the two ribosomal subunits, the mRNA to
be translated, the aminoacyl-tRNA specified by the first codon in the
message, GTP (which provides energy for the process), and initia-
tion factors that facilitate the assembly of this initiation complex (see
Figure 9.1 0). (Note: In prokaryotes, three initiation factors are known
[IF-1, IF-2, and IF-3], whereas in eukaryotes, there are over ten [des-
ignated elF to indicate eukaryotic origin]. Eukaryotes also require ATP
for initiation.) The mechanism by which the ribosome recognizes the
nucleotide sequence that initiates translation is different in eukaryotes
and prokaryotes.
In eukaryotes, the initiating AUG is recognized by a special initiator
tRNA. Recognition is facilitated by eiFs (eiF2 plus additional elF). The
amino acid-charged initiator tRNA enters the ribosomal P site, and
GTP is hydrolyzed to GOP. (Note: The initiator tRNA is the only tRNA
recognized by eiF-2, and the only tRNA to go directly to the P site.)

B. Elongation
Elongation of the polypeptide chain involves the addition of amino
acids to the carboxyl end of the growing chain. During elongation,
the ribosome moves from the 5' end to the 3' end of the mRNA that
is being translated (see Figure 9.1 0). Delivery of the aminoacyl-tRNA
whose codon appears next on the mRNA template in the ribosomal
A site is facilitated by the elongation factors (eEF-1a and eEF-1[ty).
These factors function as nucleotide exchange factors, exchanging
their GTP for the guanosine diphosphate (GOP) (from GTP hydrolysis).
The formation of the peptide bonds is catalyzed by peptldyltransfer-
ase, an activity intrinsic to the 28S rRNA found in the 60S ribosomal
subunit. Because this rRNA catalyzes the reaction, it is referred to as
a ribozyme. After the peptide bond has been formed, the ribosome
advances three nucleotides toward the 3' end of the mRNA. This pro-
cess is known as translocation and requires the participation of eEF-2
and GTP hydrolysis. This causes movement of the uncharged tRNA
into the ribosomal E site (before being released) and movement of the
peptidyl-tRNA into the P site.
V. Steps in Protein Translation 109

INITIATION

Small ribosomal unit

GUU AUG CCA CAA
----~~~~~~~~~~~-----+ 3'

Large ribosomal
GTP subunit

GDP+ P;
eiFs

Met - tRNA 1Met

P site
j __.-- Large ribosomal
subunit

E site ----
t
GUU AUG CCA__._
__.-- A site

5' _ ...................................................................... CAA

.........._-+ 3'

Psite
__.-- Large
\ ribosomal
subunit

, •.~'"NAB j)
GUU AUG CCA CAA
GTP
"-.___/
GOP

5'
E site ----

D
GUU AUG CCA - CAA
__.-- A site

3'
5' ---...J.....J--l....~.o.......J..~~----'~"--'-'--'------ 3' --....L..J'-'-'-'-J.......J..---'-...J.....J~~..__.L....J..._,____._

mRNA binding site

Small
ribosomal
subunit
Translation initiation complex
Continued on
l
next page

Figure 9.10
Steps in protein synthesis.
110 9. Translation

Amino end
ELONGATION of polypeptide

Ribosome ready fo~A /

"""' omlooacyiiRi

--r ·
~' E
'\_ site
mRNA "'\

site site

GOP

eEFs
~TP

TERMINATION

Free
polypeptide

f)[J='"
, . , ,, \j 3'

5'~
r~ , ,,
5' '
T;
Stop codon
3'

(UAG, UAA, or UGA)

a El EJ
Figure 9.10
Steps in protein synthesis- (continued).
VI. Several Antimicrobial Antibiotics Target Bacterial Translation 111

Growl"g peptide"'"'"'~

\....

Ribosomes

Figure 9.11
A polyribosome consists of several ribosomes simultaneously translating one mRNA.

C. Termination
Termination occurs when one of the three termination codons
moves into the A site (see Figure 9.1 0). Eukaryotes have a single
release factor, eRF, which recognizes all three termination codons.
The newly synthesized polypeptide may undergo further modifica-
tion as described below, and the ribosomal subunits, mRNA, tRNA,
and protein factors can be recycled and used to synthesize another
polypeptide.

D. Polysomes
Translation begins at the 5' end of the mRNA, with the ribosome
proceeding along the RNA molecule. Because of the length of most
mRNAs, more than one ribosome at a time can generally translate a
message (Figure 9.11 ). Such a complex of one mRNA and a number
of ribosomes is called a polysome or polyribosome.

E. Regulation of translation
Although gene expression is most commonly regulated at the tran-
scriptional level, the rate of protein synthesis is also sometimes reg-
ulated. An important mechanism by which this is accomplished in
eukaryotes is by covalent modification of eiF-2 (phosphorylated eiF-2
is inactive).

VI. SEVERAL ANTIMICROBIAL ANTIBIOTICS TARGET

BACTERIAL TRANSLATION

The initiation process in protein synthesis is different in prokaryotes

and eukaryotes as shown in Table 9.1. Many antibiotics that are used
to combat bacterial infections in humans take advantage of the differ-
ences between the mechanisms for protein synthesis in prokaryotes and
eukaryotes (Table 9.2).
112 9. Translation

Table 9.1 : Differences Between Prokaryotes and Eukaryotes

in the Initiation of Protein Synthesis

Eukaryotes Prokaryotes
Binding of mRNA to Cap at 5'-end of mRNA A specific sequence
small ribosomal binds to eiFs and 40S upstream of Initiating AUG
subunit ribosomal subunit. mRNA binds to complementary
is scanned for first AUG sequence In 16S RNA
First amino acid Methionine Formyl methionine
Initiation factors eiFs (12 or more) IFs (3)
Ribosomes 80S (40S and 60S subunits) 70S (30S and 50S subunits)

VII. POSTTRANSLATIONAL MODIFICATION OF

POLYPEPTIDE CHAINS

Many polypeptide chains are covalently modified, either while they are
still attached to the ribosome or after their synthesis has been completed.
Because the modifications occur after the translation is initiated, they are
called posttranslational modifications. These modifications may include
the removal of a part of the translated sequence or the covalent addition
of one or more chemical groups required for protein activity. Some types
of posttranslational modifications are listed below.

A. Trimming
Many proteins destined for secretion from the cell are initially made
as large, precursor molecules that are not functionally active. Portions
of the protein chain must be removed by specialized endoproteases,
resulting in the release of an active molecule. The cellular site of the
cleavage reaction depends on the protein to be modified. For example,
some precursor proteins are cleaved in the endoplasmic reticulum or
the Golgi apparatus, others are cleaved in developing secretory ves-
icles, and still others, such as collagen, are cleaved after secretion.
Zymogens are inactive precursors of secreted enzymes (including
the proteases required for digestion). They become activated through
cleavage when they reach their proper sites of action. For example,
the pancreatic zymogen, trypsinogen, becomes activated to trypsin
in the small intestine.

lu The synthesis of enzymes as zymogens protects the cell

from being digested by its own products.

Table 9.2: Effects of Antibiotics on Prokaryotic Protein

Synthesis
Streptomycin Inhibits initiation and causes misreading
Tetracycline Binds to the 30S subunit and inhibits the binding of
aminoacyl-tRNAs
Erythromycin Binds to the 50S subunit and inhibits translocation
VII. Posttranslational Modification of Polypeptide Chains 113

B. Covalent alterations
Proteins, both enzymatic and structural, may be activated or inac-
tivated by the covalent attachment of a variety of chemical groups.
Examples of these modifications include (Figure 9.12):

[ Phosphorylation } [ Hydroxylation l
II
Phosphate H
I
0
II
C=O
I
-..fVVVVVV'- HN -
I
C - CO -JVVVV
I
II
-o- P- O - CH2 - CH
II
0
I I
NH Hy~roxyl~ yH
H2C,
,CH2

res1due OH
Serine ~
Protein
[ Carboxylation J
Mature clotting
H factors

VVV'-N/-~~TAA]ftA%•:::~
'\. (Gia) residue
coo- coo-

[ Glycosylation ) l Biotinylated enzyme J

HO~~
H

L
VVVV'- N- CH- C -'VVVV
1 II
CH2 0
Lysyl residue CH
2
of carboxylase •
~- CH2 - ?H ...--- Serine
enzyme :......---- yH2
CH2
'

··s:rl
NH NH
I
C= O
I
N-Acetyl-
CHs

galactosamine s;ouo

Biotin-enzyme

II

Farnesyl group

Figure 9.12
Posttranslational modifications of some amino acid residues.
114 9. Translation

1. Phosphorylation: Phosphorylation occurs on the hydroxyl groups

of serine, threonine, or, less frequently, tyrosine residues in a pro-
tein. This phosphorylation is catalyzed by one of a family of pro-
tein kinases and may be reversed by the action of cellular protein
phosphatases. The phosphorylation may increase or decrease the
functional activity of the protein.

2. Glycosylation: Many of the proteins that are destined to become

part of a plasma membrane or lysosome, or to be secreted from
the cell, have carbohydrate chains attached to serine or threonine
hydroxyl groups (C-linked) or the amide nitrogen of asparagine
(N-Iinked). The addition of sugars occurs in the endoplasmic retic-
ulum and the Golgi apparatus. Sometimes, glycosylation is used
to target the proteins to specific organelles. For example, enzymes
destined to be incorporated into lysosomes are modified by the
phosphorylation of mannose residues (see Chapter 11 ).

3. Hydroxylation: Proline and lysine residues of the a chains of col-

lagen are extensively hydroxylated in the endoplasmic reticulum.

4. Other covalent modifications: These may be required for the

functional activity of a protein. For example, additional carboxyl
groups can be added to glutamate residues by vitamin K-depen-
dent carboxylation. The resulting y-carboxyglutamate residues are
essential for the activity of several of the blood-clotting proteins.
Biotin is covalently bound to the £-amino groups of lysine residues
of biotin-dependent enzymes that catalyze carboxylation reac-
tions, such as pyruvate carboxylase. Attachment of lipids, such
as farnesyl groups, can help anchor proteins in membranes. In
addition, many proteins are acetylated after translation.

Chapter Summary

• Codons are composed of three nucleotide bases presented in the mRNA language of A, G, C, and U. There are 64 pos-
sible combinations with 61 coding for the 20 common amino acids and three termination signals.
• The genetic code is specific, universal, degenerate, nonoverlapping, and commaless.
• Mutations are a result of altering the nucleotide sequence.
• Requirements for protein synthesis include all the amino acids that eventually appear in the finished protein, at least
one specific type of tRNA for each amino acid, one aminoacyl-tRNA synthetase for each amino acid, the mRNA coding
for the protein to be synthesized, ribosomes, protein factors, and ATP and GTP as energy sources.
• The formation of the peptide bond is catalyzed by peptidyl transferase, an activity intrinsic to the large ribosomal subunit.
• More than one ribosome at a time can translate a message forming a polysome.
• Numerous antibiotics interfere with the process of protein synthesis selectively in prokaryotes and eukaryotes.
• Many polypeptides are covalently modified after synthesis.