POST TRANSLATIONAL

MODIFICATION(PTM)
S.HAMSHAVARSHINI

WHAT IS PTM?
ADDITION OF GROUPS OR DELETION OF PARTS TO MAKE A
FINISHED PROTEIN.

WHAT ARE THE GROUPS??

-METYHL
-ACETYL
-GLYCO
-PHOSPHO

PURPOSE OF PTM
QUALITY CONTROL -chaperons, glycosylation
DEGRADATION OF MISFOLDED PROTEIN ubiqutination
PROPER PROTEIN FUNCTION glycosylation ,
phosphorylation, ubiqutination
TARGET PROTEIN TO PROPER LOCATION -acylation

Post translational MODIFICATIONS

1. Methylation
2. Acylation
3. Ubiquitination
4. Proteolysis
5. Sulfation
6. Lipidation
7. Vitamin C dependent modification
8. Vitamin k dependent modification
9. Phosphorylation
10.Glycosylation

1. METHYLATION
Post-translational methylation of proteins occurs on nitrogen and
oxygen.
The activated methyl donor is S-adenosthionineylme (SAM).
Nitrogen methylations are found on the imidazole ring of histidine ,
the R-group amides of glutamate and aspartate and the -amine of
lysine residues .
Methylation of the oxygen of the R-group carboxylates of gutamate
and aspartate also takes place and forms methyl esters.
The most common methylations are on the -amine of lysine
residues. Methylation of lysine residues in histones in DNA is an
important regulator of chromatin structure and consequently of
transcriptional activity.

2. ACYLATION
Many proteins are modified at their N-termini following
synthesis.
In most cases the initiator methionine is hydrolyzed and
an acetyl group is added to the new N-terminal amino
acid.
Acetyl-CoA is the acetyl donor for these reactions.
chromatinproteins and metabolic enzymes are highly
represented, indicating that acylation has a
considerable impact ongene expression
andmetabolism.

3. UBIQUITINATION
Degradation of proteins in the proteasome occurs via an
ATP-dependent mechanism.
In eukaryotic cells the proteasome is found in the
cytosol and the nucleus and has a large mass such that
it has a sedimentation coefficient of 26S. The 26S
proteasome comprises a 20S barrel-shaped catalytic
core as well as 19S regulatory complexes at both ends.
Proteins that are to be degraded by the proteasome are
first tagged by attachment of multimers of the 76 amino
acid polypeptide ubiquitin.

Process of ubiquitination and proteasome-mediated protein degradation

4. PROTEOLYSIS
Most proteins undergo proteolytic cleavage following
translation.
The simplest form of this is the removal of the initiation
methionine.
Many proteins are synthesized as inactive precursors
that are activated under proper physiological conditions
by limited proteolysis.
Example of a preproprotein are insulin and zymogens.

5. SULFATION
Sulfate modification of proteins occurs at tyrosine
residues.
Tyrosine sulfation is accomplished via the activity of
tyrosyl protein sulfotransferases (TPST) which are
membrane associated enzymes of the trans-Golgi
network.
The universal sulfate donor for these TPST enzymes is
3'-phosphoadenosyl-5'-phosphosulphate (PAPS).
Addition of sulfate occurs almost exclusively on
secreted and trans-membrane spanning proteins.

 The addition of sulfate to tyrosine is believed to play a

role in the modulation of protein-protein interactions of
secreted and membrane bound proteins.
The process of tyrosine sulfation has been shown to be
critical for the processes of various immune functions,
intracellular trafficking, and ligand recognition by
several G-protein-coupled receptors (GPCRs).

6. LIPIDATION
PRENYLATION
Prenylation refers to the addition of the 15 carbon farnesyl group or
the 20 carbon geranylgeranyl group to acceptor proteins, both of
which are isoprenoid compounds derived from the cholesterol
biosynthetic pathway.
The isoprenoid groups are attached to cysteine residues at the
carboxy terminus of proteins in a thioether linkage (C-S-C).
CAAX, where C is cysteine, A is any aliphatic amino acid (except
alanine) and X is the C-terminal amino acid.
In order for the prenylation reaction to occur the three C-terminal
amino acids (AAX) are first removed. Following attachment of the
prenyl group the carboxylate of the cysteine is methylated in a
reaction utilizing S-adenosylmethionine as the methyl donor.
RAS-related G-proteins.

Prenylation

MYRISTOYLATION
Some proteins have the 14 carbon myristoyl group added to their Ntermini. The donor for this modification is myristoyl-CoA. This modification
allows association of the modified protein with membranes. The catalytic
subunit of cyclicAMP-dependent protein kinase (PKA) is myristoylated.
PALMITOYLATION
S-palmitoylation adds a C16 palmitoyl group from palmitoyl-CoA to the
thiolate side chain of cysteine residues via palmitoyl acyl transferases
(PATs).This anchor can permanently anchor the protein to the membrane.
This localization can be reversed, though, by thioesterases that break the
link between the protein and the anchor; thus, S-palmitoylation is used as
an on/off switch to regulate membrane localization. S-palmitoylation is
often used to strengthen other types of lipidation, such as myristoylation
or farnesylation.

GPI anchors
GPI anchors tether cell surface proteins to the plasma
membrane. These hydrophobic moieties are prepared in
the ER, where they are then added to the nascent
protein. GPI-anchored proteins are often localized to
cholesterol- and sphingolipid-rich lipid rafts, which act
as signaling platforms on the plasma membrane.

7.VITAMIN C-dependent modification

Modifications of proteins that depend upon vitamin C as
a cofactor include proline and lysine hydroxylations and
carboxy terminal amidation.
The hydroxylating enzymes are identified as prolyl
hydroxylase and lysyl hydroxylase.
The donor of the amide for C-terminal amidation is
glycine.
hydroxylated proteins are the collagens.
Several peptide hormones such as oxytocin and
vasopressin have C-terminal amidation.

8. VITAMIN K-dependent
modification
Vitamin K is a cofactor in the carboxylation of glutamic
acid residues catalyzed by the enzyme gamma-glutamyl
carboxylase (-glutamyl carboxylase).
The result of this type of reaction is the formation of a
-carboxy glutamate (gamma-carboxy glutamate),
referred to as a gla residue.
The reaction catalyzed by -glutamyl carboxylase is the
one that incorporates the gla-residue but two additional
enzyme activities are required to convert vitamin K back
to its active hydroquinone (quinol) form.

 The incorpration of a gla-residue into a protein such as

prothrombin requires the hydroquinone (KH2) form of
vitamin K (either K1, K2, or synthetic K3).
The utilization and regeneration of the KH2 form in the
overall process of the -glutamyl carboxylase (GGCX)
reaction is referred to as the vitamin K cycle.
Either following the carboxylation, or directly from
dietary quinone forms of vitamin K, the action of vitamin
K epoxide reductase (VKORC1) is to provide a continuous
source of the KH2 form.

References
Themedicalbiochemistrypage.org, 19962013.
Post translational modifiers, Anthony Francisco and
Nicole Benson.
Modification-specific proteomics: Characterization of
posttranslational modifications by mass spectrometry,
Jensen O. N. 2004.
PREMIER Biosoft, 1994-2014 .