BIOCHEMICAL GENETICS
Dr. Noor-ul-Ain Waheed
Assistant Professor,
Dept. of Biochemistry, KEMU
• Post-translational modifications
of newly synthesized protein
• Protein targeting
Coupling of transcription and translation in
bacteria
Post translational Modifications
• Many proteins are modified after translation
Modification of Individual Amino Acids
• The hydroxyl groups of
certain residues of
some proteins are
enzymatically
phosphorylated by ATP
• Significance
• milk protein casein has
many phosphoserine
groups that bind Ca2+
• Regulate the activity of
many enzymes and
regulatory proteins
Addition of Isoprenyl Groups
• Eukaryotic proteins are modified by the addition of groups derived
from isoprene (isoprenyl groups).
• A thioether bond is formed between the isoprenyl group and a Cys
residue of the protein.
• The isoprenyl group helps to anchor the protein in a membrane.
Formation of Disulfide Cross-Links
• After folding into their native conformations,
some proteins form intrachain or interchain
disulfide bridges between Cys residues.
• In eukaryotes, disulfide bonds are common in
proteins to be exported from cells.
• The cross-links formed in this way help to
protect from denaturation in the extracellular
environment.
Addition of Prosthetic Groups
• Many proteins require for their activity
covalently bound prosthetic groups.
Two examples are :
1. The biotin molecule of acetyl-CoA
carboxylase
2. The heme group of hemoglobin or
cytochrome c.
Protein Targeting
1. The targeting pathway begins with initiation of protein synthesis on free
ribosomes.
2. The signal sequence appears early in the synthetic process, because it is at
the amino terminus, which as we have seen is synthesized first.
3. As it emerges from the ribosome, the signal sequence—and the ribosome
itself—is bound by the large signal recognition particle (SRP); SRP then
binds GTP and halts elongation of the polypeptide when it is about 70
amino acids long and the signal sequence has completely emerged from
the ribosome.
4. The GTP-bound SRP now directs the ribosome (still bound to the mRNA)
and the incomplete polypeptide to GTP-bound SRP
receptors in the cytosolic face of the ER; the nascent polypeptide is
delivered to a peptide translocation complex in the ER, which may interact
directly with the ribosome.
5. SRP dissociates from the ribosome, accompanied by hydrolysis of GTP in
both SRP and the SRP receptor.
6. Elongation of the polypeptide now resumes, with the ATP-driven
translocation complex feeding the growing polypeptide into the ER lumen
until the complete protein has been synthesized.
7. The signal sequence is removed by a signal peptidase within the ER lumen;
8. the ribosome dissociates &
9. is recycled.
Phosphorylation of mannose residues
on lysosome-targeted enzymes
Glycosylation Plays a Key Role
in Protein Targeting
Signal Sequences for Nuclear
Transport Are Not Cleaved
• The signal sequence that targets a protein to the
nucleus—the nuclear localization sequence
(NLS)—is not removed after the protein arrives
at its destination.
• NLS contains four to eight amino acid residues
and include several consecutive basic (Arg or
Lys) residues.
Regulation of gene
expression in prokaryotes
The lac Operon of E. coli
The lac operon of E. coli is a segment of DNA
that includes a promoter, an operator, and the
three structural genes that code for lactose-
metabolizing enzymes.
How to control working of
repressors?
• Two regulatory mechanisms
• Induction (an inducer inactivates the
repressor)
• Repression (a co-repressor is required to
activate the repressor)
corepressor
•Regulation of lac
operon & trp
operon expression
in [Link] bacteria
• What happens to the
expression of the lac
operon when both
glucose and lactose are
present?
Positive regulation of
the lac operon by CRP
Regulation of gene
transcription
conserves energy in
[Link]
Transcriptional
attenuation when trp ↑
Transcriptional
attenuation when trp ↓
ribosome
Difference between prokaryotic & eukaryotic gene
expression regulation
Thanks