3 Step Elongation Phase

 Elongation is a cycle of events

 Step 1 – aminoacyl-tRNA comes into empty A-site next to the
occupied P-site; pairs with the codon
 Step 2 – C’ end of peptide chain uncouples from tRNA in P-site
and links to AA in A-site
 Peptidyl transferase responsible for bond formation
 Each AA added carries the energy for the addition of the
next AA
 Step 3 – peptidyl-tRNA moves to the P-site; requires hydrolysis
of GTP
 tRNA released back to the cytoplasmic pool

Initiation Process
 Determines whether mRNA is synthesized and sets the reading
frame that is used to make
the protein
 Initiation process brings the
ribosomal subunits together
at the site where the peptide
should begin
 Initiator tRNA brings in Met
 Initiator tRNA is
different than the tRNA
that adds other Met

Ribosomal Assembly Initiation

Phase
 Initiation factors (IFs)
catalyze the steps – not well
defined
 Step 1 – small ribosomal
subunit with the IF finds the
start codon –AUG
 Moves 5’ to 3’ on
mRNA
 Initiator tRNA brings in the 1st AA which is always
Met and then can bind the mRNA
 Step 2 – IF leaves and then large subunit can bind –
protein synthesis continues
 Met is at the start of every protein until post-
translational modification takes place

Eukaryotic vs Procaryotic

 Procaryotic
 No CAP; have specific ribosome binding site
upstream of AUG
 Polycistronic – multiple proteins from same mRNA
 Eucaryotic
 Monocistronic – one polypeptide per mRNA
Protein Release
 Protein released when a STOP codon is encountered
 UAG, UAA, UGA (must know these sequences!)
 Cytoplasmic release factors bind to the stop codon that gets to
the A-site; alters the peptidyl transferase and adds H2O instead
of an AA
 Protein released and the ribosome breaks into the 2 subunits
to move on to another mRNA

Polyribosomes
 As the ribosome moves down the mRNA, it allows for the
addition of another ribosome and the start of another protein
 Each mRNA has multiple ribosomes attached, polyribosome or
polysome
Regulation of Protein Synthesis
 Lifespan of proteins vary, need method to remove old or
damaged proteins
 Enzymes that degrade proteins are called proteases –
process is called proteolysis
 In the cytosol there are large complexes of proteolytic
enzymes that remove damaged proteins
 Ubiquitin, small protein, is added as a tag for disposal of
protein

Protein Synthesis
 Protein synthesis takes the most energy input of all the
biosynthetic pathways
 4 high-energy bonds required for each AA addition
 2 in charging the tRNA (adding AA)
 2 in ribosomal activities (step 1 and step 3 of elongation phase)