3. Turnover of protein is NOT constant Half lives of proteins vary from minutes to
infinity “Normal” proteins – 100-200 hrs
Short-lived proteins-regulatory proteins enzymes that catalyze committed steps
transcription factors
Long-lived proteins-Special cases (dentin, crystallins)
3/11/2018
Sweta kumari,16BBT0124,CELL BIOLOGY AND
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4. Proteins are not degraded at the same rate :-
ENZYME Half-life
Ornithine decarboxylase 11 minutes
delta-Aminolevulinate synthetase 70 minutes
Catalase 1.4 days
Tyrosine aminotransferase 1.5 hours
Tryptophan oxygenase 2 hours
Glucokinase 1.2 days
Lactic dehydrogenase 16 days
3/11/2018Sweta kumari,16BBT0124,CELL BIOLOGY AND BIOCHEMISTRY
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5. May depend on tissue distribution:
Example: Lactic Acid Dehydrogenase
Tissue Half-life
Heart -1.6 days
Muscle - 31 days
Liver -16 days
Protein degradation is a regulated process:
Example: Acetyl CoA carboxylase
Nutritional state
Half-life Fed-48 hours
Fasted -18 hours
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Sweta kumari,16BBT0124,CELL BIOLOGY AND
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6. Ubiquitin/Proteasome Pathway:
80-90% Most intracellular proteins
Lysosomal processes:
10-20% Extracellular proteins
Cell organelles
Some intracellular proteins
Proteasomes:
Large (26S) multiprotein complex (28 subunits)
Degrades ubiquitinated proteins
Lysosomes:
Basal degradation – non-selective
Degradation under starvation – selective for “KFERQ” proteins
3/11/2018Sweta kumari,16BBT0124,CELL BIOLOGY AND BIOCHEMISTRY
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7. The Ubiquitin/Proteasome PATHWAY:
Small peptide that is a “TAG”
76 amino acids
C-terminal glycine - isopeptide bond with the e-amino group of lysine
residues on the substrate
Attached as monoubiquitin or polyubiquitin chains
Three genes in humans: Two are stress genes (B and C) One, UbA as a
fusion protein
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Sweta kumari,16BBT0124,CELL BIOLOGY AND
BIOCHEMISTRY
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9. Four Main Steps:
UBIQUITINATION
RECOGNITION
DEGRADATION
DEUBIQUITINATION
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Sweta kumari,16BBT0124,CELL BIOLOGY AND
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10. UBIQUITINATION:
First, Ubiquitin is activated by forming a link to “enzyme 1” (E1).
Then, ubiquitin is transferred to one of several types of “enzyme 2” (E2).
Then, “enzyme 3” (E3) catalizes the transfer of ubiquitin from E2 to a Lys e-amino
group of the “condemned” protein.
Lastly, molecules of Ubiquitin are commonly conjugated to the protein to be
degraded by E3s & E4s
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Sweta kumari,16BBT0124,CELL BIOLOGY AND
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11. Ubiquitinated proteins are degraded by the proteasome:
Ubiquitinated proteins are degraded in the cytoplasm and nucleus by the
proteasome.
Proteasomal protein degradation consumes ATP.
The proteasome degrades the proteins to ~8 amino-acid peptides.
Access of proteins into the proteasome is tightly regulated.
The peptides resulting from the proteasome activity diffuse out of the proteasome
freely.
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Sweta kumari,16BBT0124,CELL BIOLOGY AND
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13. LYSOSOMES:
Digestive System of the Cell
Digests
– ingested materials
– obsolete cell components
• Degrades macromolecules of all types
– Proteins
– Nucleic acids
– Carbohydrates
– Lipids
• Heterogeneous
3/11/2018Sweta kumari,16BBT0124,CELL BIOLOGY AND BIOCHEMISTRY
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14. Protein degradation in the lysosomes
Lysosomes degrade extracellular proteins that the cell incorporates by
endocytosis.
Lysosomes can also degrade intracellular proteins that are enclosed in other
membrane-limited organellas.
In well-nourished cells, lysosomal protein degradation is non-selective (non-
regulated).
In starved cells, lysosomes degrade preferentially proteins containing a KFERQ
“signal” peptide.
The regression of the uterus after childbirth is mediated largely by lysosomal
protein degradation
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Sweta kumari,16BBT0124,CELL BIOLOGY AND
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18. Conclusions and future perspectives:
Only a few targeting signals have been identified, and the mechanisms that
underlie the regulation of the system are still largely unknown?
While the system has been implicated in the pathogenesis of several diseases, the
underlying mechanisms, as well as its potential involvement in many other
diseases, are still an enigma?
Why are there so many ubiquitinating enzymes if prior modifications such as
phosphorylation or damage are triggering events?
Do DUBs show substrate specificity, perhaps by regulating the levels of
ubiquitination of specific subsets of proteins?
What are the binding sites for polyubiquitin chains on the microtubules and on
the proteasome itself?
3/11/2018
Sweta kumari,16BBT0124,CELL BIOLOGY AND
BIOCHEMISTRY
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