In the final stage of protein synthesis, the nascent polypeptide chain is folded and processed into its biologically

active form. During or after its synthesis, the polypeptide progressively assumes its native conformation, with
the formation of appropriate hydrogen bonds and van der Waals, ionic, and hydrophobic interactions. Some
newly made proteins, both prokaryotic and eukaryotic, do not attain their final biologically active conformation
until they have been altered by one or more processing reactions called posttranslational modifications.
Amino-Terminal and Carboxyl-Terminal Modifications
The first residue inserted in all polypeptides is N-formylmethionine in bacteria! or methionine in eukaryotes!.
"owever, the formyl group, the amino-terminal #et residue, and often additional amino-terminal and, in some
cases, carbo$yl-terminal! residues may be removed en%ymatically in formation of the final functional protein.
In as many as &'( of eukaryotic proteins, the amino group of the amino-terminal residue is N-acetylated after
translation. )arbo$yl-terminal residues are also sometimes modified.
Loss of Signal Sequences
The *& to +' residues at the amino-terminal end of some proteins play a role in directing the protein to its
ultimate destination in the cell. Such signal sequences are ultimately removed by specific peptidases.
Modification of Individual Amino Acids
The hydro$yl groups of certain Ser, Thr, and Tyr residues of some proteins are en%ymatically phosphorylated by
,T-. the phosphate groups add negative charges to these polypeptides. The functional significance of this
modification varies from one protein to the ne$t. /or e$ample, the milk protein casein has many phosphoserine
groups that bind )a
0
. ,nd as we have seen in numerous instances, phosphorylation dephosphorylation cycles
regulate the activity of many en%ymes and regulatory proteins.
1$tra carbo$yl groups may be added to 2lu residues of some proteins. /or e$ample, the blood-clotting protein
prothrombin contains a number of -carbo$yglutamate residues in its amino-terminal region, introduced by an
en%yme that re3uires vitamin 4.
.
Attachment of Carbohydrate Side Chains
The carbohydrate side chains of glycoproteins are attached covalently during or after synthesis of the
polypeptide. In some glycoproteins, the carbohydrate side chain is attached en%ymatically to ,sn residues N-
linked oligosaccharides!, in others to Ser or Thr residues O-linked oligosaccharides!. #any proteins that
function e$tracellularly, as well as the lubricating proteoglycans that coat mucous membranes, contain
oligosaccharide side chains.
Addition of Isoprenyl roups
, number of eukaryotic proteins are modified by the addition of groups derived from isoprene isoprenyl
groups!. , thioether bond is formed between the isoprenyl group and a )ys residue of the protein. The isoprenyl
groups are derived from pyrophosphorylated intermediates of the cholesterol biosynthetic pathway, such as
farnesyl pyrophosphate. -roteins modified in this way include the 5as proteins, products of the ras oncogenes
and proto-oncogenes, and 2 proteins, and lamins, proteins found in the nuclear matri$. The isoprenyl group
helps to anchor the protein in a membrane. The transforming carcinogenic! activity of the ras oncogene is lost
when isoprenylation of the 5as protein is blocked, a finding that has stimulated interest in identifying inhibitors
of this posttranslational modification pathway for use in cancer chemotherapy.
Addition of !rosthetic roups
#any prokaryotic and eukaryotic proteins re3uire for their activity covalently bound prosthetic groups. Two
e$amples are the biotinmolecule of acetyl-)o, carbo$ylase and the heme group of hemoglobin or cytochrome
c.
!roteolytic !rocessing
#any proteins are initially synthesi%ed as large, inactive precursor polypeptides that are proteolytically trimmed
to form their smaller, active forms. 1$amples include proinsulin, some viral proteins, and proteases such as
chymotrypsinogen and trypsinogen
"ormation of #isulfide Cross-Lin$s
,fter folding into their native conformations, some proteins form intrachain or interchain disulfide bridges
between )ys residues. In eukaryotes, disulfide bonds are common in proteins to be e$ported from cells. The
cross-links formed in this way help to protect the native conformation of the protein molecule from
denaturation in the e$tracellular environment, which can differ greatly from intracellular conditions and is
generally o$idi%ing